Study on the reversal of epileptic drug resistance by tetramethylpyrazine in combination with carbamazepine through modulation of P-Glycoprotein expression in rat brain microvessel endothelial cell.

The purpose of this study was to detect the changes of P-Glycoprotein (P-GP) expression in rat brain microvessel endothelial cell line RBE4 after the action of Tetramethylpyrazine (TMP) on Carbamazepine (CBZ), so as to clarify the potential mechanism of TMP combined with CBZ against intractable epilepsy drug resistance. The RBE4 cell line was utilized for in vitro analysis. Cells were divided into control, CBZ, and CBZ-TMP group. The expression of P-GP was assessed using Western blot and reverse transcription polymerase chain reaction (RT-PCR). Intracellular concentration of CBZ was measured through high-performance liquid chromatography (HPLC). The differential expression of mRNA was evaluated by RNA sequencing. The intracellular concentration of CBZ in the CBZ-TMP group was significantly higher than that in other groups. The expression of P-GP in the CBZ group was significantly higher than that in the control group, while in the CBZ&TMP group, it was significantly lower than that in the other groups. Comparative analysis also revealed some differentially expressed genes. Compared with the CBZ group, FAM106A, SLC3A2, TENM2, etc. were upregulated most significantly in the CBZ&TMP group. ZBTB10, WDR7, STARD13, etc. were downregulated most significantly. Results suggest that TMP increases the intracellular concentration of CBZ, downregulates the expression of P-GP increased by CBZ, and modulates related cellular metabolism and signaling pathways, thus reversing the drug resistance mechanism of intractable epilepsy, providing a theoretical basis for the combination of traditional Chinese medicine and antiepileptic drugs.

Mesenchymal Stem Cells Combined With Electroacupuncture Treatment Regulate the Subpopulation of Macrophages and Astrocytes to Facilitate Axonal Regeneration in Transected Spinal Cord.

OBJECTIVE: Herein, we investigated whether mesenchymal stem cells (MSCs) transplantation combined with electroacupuncture (EA) treatment could decrease the proportion of proinflammatory microglia/macrophages and neurotoxic A1 reactive astrocytes and inhibit glial scar formation to enhance axonal regeneration after spinal cord injury (SCI). METHODS: Adult rats were divided into 5 groups after complete transection of the spinal cord at the T10 level: a control group, a nonacupoint EA (NA-EA) group, an EA group, an MSC group, and an MSCs+EA group. Immunofluorescence labeling, quantitative real-time polymerase chain reaction, enzyme-linked immunosorbent assay, and Western blots were performed. RESULTS: The results showed that MSCs+EA treatment reduced the proportion of proinflammatory M1 subtype microglia/macrophages, but increased the differentiation of anti-inflammatory M2 phenotype cells, thereby suppressing the mRNA and protein expression of proinflammatory cytokines (tumor necrosis factor-α and IL-1ß) and increasing the expression of an anti-inflammatory cytokine (interleukin [IL]-10) on days 7 and 14 after SCI. The changes in expression correlated with the attenuated neurotoxic A1 reactive astrocytes and glial scar, which in turn facilitated the axonal regeneration of the injured spinal cord. In vitro, the proinflammatory cytokines increased the level of proliferation of astrocytes and increased the expression levels of C3, glial fibrillary acidic protein, and chondroitin sulfate proteoglycan. These effects were blocked by administering inhibitors of ErbB1 and signal transducer and activator of transcription 3 (STAT3) (AG1478 and AG490) and IL-10. CONCLUSION: These findings showed that MSCs+EA treatment synergistically regulated the microglia/macrophage subpopulation to reduce inflammation, the formation of neurotoxic A1 astrocytes, and glial scars. This was achieved by downregulating the ErbB1-STAT3 signal pathway, thereby providing a favorable microenvironment conducive to axonal regeneration after SCI.

Governor Vessel Electro-Acupuncture Promotes the Intrinsic Growth Ability of Spinal Neurons through Activating Calcitonin Gene-Related Peptide/α-Calcium/Calmodulin-Dependent Protein Kinase/Neurotrophin-3 Pathway after Spinal Cord Injury.

Spinal cord injury (SCI) invariably results in neuronal death and failure of axonal regeneration. This is attributed mainly to the hostile microenvironment and the poor intrinsic regrowth capacity of the injured spinal neurons. We have reported previously that electro-acupuncture on Governor Vessel acupoints (GV-EA) can promote neuronal survival and axonal regeneration of injured spinal cord. However, the underlying mechanism for this has remained uncertain. The present study aimed to explore the neural afferent pathway of GV-EA stimulation and the possible mechanism by which GV-EA can activate the intrinsic growth ability of injured spinal neurons. By cholera toxin B (CTB) retrograde labeling, immunostaining, and enzyme-linked immunosorbent assay (ELISA), we showed here that GV-EA could stimulate the spinal nerve branches of the dorsal root ganglion cells. This would then increase the release of calcitonin gene-related peptide (CGRP) from the afferent terminals in the spinal cord. It is of note that the effect was abrogated after dorsal rhizotomy. Additionally, both in vivo and in vitro results showed that CGRP would act on the post-synaptic spinal cord neurons and triggered the synthesis and secretion of neurotrophin-3 (NT-3) by activating the calcitonin gene-related peptide (CGRP)/ receptor activity-modifying protein (RAMP)1/calcium/calmodulin-dependent protein kinase (αCaMKII) pathway. Remarkably, the observed effect was prevented by the dorsal rhizotomy and the blockers of the CGRP/RAMP1/αCaMKII pathway. More importantly, increase in NT-3 promoted the survival, axonal regrowth, and synaptic maintenance of spinal cord neurons in the injured spinal cord. Therefore, it is concluded that increase in NT-3 production is one of the mechanisms by which GV-EA can activate the intrinsic growth ability of spinal neurons after SCI. The experimental results have reinforced the theoretical basis of GV-EA for its clinical efficacy in patients with SCI.

A Retrospective Study of Gemcitabine and Carboplatin With or Without Intravenous Vitamin C on Patients With Advanced Triple-Negative Breast Cancer.

Background: This is a retrospective study to examine the effect of chemotherapy with or without intravenous vitamin C (IVC) on women with advanced triple-negative breast cancer (TNBC). Methods: From 2008 to 2016, a total of 113 patients with pathologically confirmed TNBC at Clifford Hospital were evaluated, and 70 patients were matched and divided into IVC (treatment group) and non-IVC groups (control group). The match was according to age, menopausal status, and metastatic sites. In the control group, 35 patients received gemcitabine and carboplatin. In the treatment group, 35 patients received the same chemotherapy plus IVC. Results: Baseline characteristics were not significantly different between the 2 groups. According to the criteria of RECIST 1.1 (Response Evaluation Criteria in Solid Tumors), enhanced computed tomography scan was compared after 2 cycles of chemotherapy. In the treatment group, 2/35 cases had a complete remission (CR), 15/35 cases had partial remission (PR), and 13/35 cases had stable disease (SD). The response rate was 48.6%. In the control group, there were no CR cases, 14/35 cases had PR, 14/35 cases had SD, and the response rate was 40.0% (P > .05). The median progression-free survival time and median overall survival time was 7 months (95% confidence interval [CI] =1.5-28.5 months) and 27 months (95% CI = 4-40 months) in the treatment group compared with 4.5 months (95% CI = 1.5-8 months) and 18 months (95% CI = 3-26 months) in the control group (P < .05). All patients experienced diverse reactions in the gastrointestinal tract and myelosuppression. The incidence of adverse reactions in the treatment group was significantly lower than that of the control group (P < .05). Conclusion: IVC may have an effect on improving the prognosis of patients with advanced TNBC.

[Post-stroke hand spasm treated with penetrating acupuncture combined with kinesiotherapy: a randomized controlled trial].

OBJECTIVE: To compare the differences in the clinical effect on post-stroke hand spasm among the combined treatment of penetrating acupuncture and kinesiotherapy, the simple application of penetrating acupuncture and the simple application of kinesiotherapy. METHODS: A total of 105 patients with post-stroke hand spasm were randomized into a penetrating acupuncture group, a kinesiotherapy group and a combined treatment group, 35 cases in each one, of which, 2 cases were dropped out in either the combined treatment group and the penetrating acupuncture group, and 1 case dropped out in the kinesiotherapy group. The routine rehabilitation training, e.g. occupational therapy and Bobath exercise and medication were adopted in all of the three groups. In the penetrating acupuncture group, the penetrating needling technique was exerted from Hegu (LI 4) to Houxi (SI 3) and from Waiguan (TE 5) to Sidu (TE 9) on the affected side. In the kinesiotherapy group, the persistent movement or passive movement was exerted on the wrist joint, the metacarpophalangeal joints and the interphalangeal joints. In the combined treatment group, the penetrating acupuncture (the same as the penetrating acupuncture group) was exerted combined with kinesiotherapy (the same as the kinesiotherapy group). In each group, the treatment was given once a day, 30 min in each time, 6 treatments a week in total, with the interval of 1 day between the courses. The treatment for 2 weeks was as one course and 2 courses were required totally. Before and after treatment, the scores of hand spasm index, hand-wrist motor function and the activity of daily living (ADL) were compared in each group. RESULTS: After treatment, the scores of hand spasm index were reduced as compared with those before treatment in each group (P<0.05) and the scores of hand-wrist motor function and ADL were increased significantly as compared with those before treatment in each group (P<0.05). After treatment, the reducing degree of the score of hand spasm index in the combined treatment group was greater than the penetrating acupuncture group and the kinesiotherapy group (P<0.01), and the increasing degree of the scores of hand-wrist motor function and ADL were higher than either the penetrating acupuncture group or the kinesiotherapy group (P<0.01). The improvements in each index were not different statistically between the kinesiotherapy group and the penetrating acupuncture group (P>0.05). CONCLUSION: Compared with the simple application of either penetrating acupuncture or kinesiotherapy, the combined treatment of them achieves the significant improvements in hand spasm degree, hand wrist motor function and ADL in patients with stroke.

Electroacupuncture Facilitates the Integration of Neural Stem Cell-Derived Neural Network with Transected Rat Spinal Cord.

The hostile environment of an injured spinal cord makes it challenging to achieve higher viability in a grafted tissue-engineered neural network used to reconstruct the spinal cord circuit. Here, we investigate whether cell survival and synaptic transmission within an NT-3 and TRKC gene-overexpressing neural stem cell-derived neural network scaffold (NN) transplanted into transected spinal cord could be promoted by electroacupuncture (EA) through improving the microenvironment. Our results showed that EA facilitated the cell survival, neuronal differentiation, and synapse formation of a transplanted NN. Pseudorabies virus tracing demonstrated that EA strengthened synaptic integration of the transplanted NN with the host neural circuit. The combination therapy also promoted axonal regeneration, spinal conductivity, and functional recovery. The findings highlight EA as a potential and safe supplementary therapeutic strategy to reinforce the survival and synaptogenesis of a transplanted NN as a neuronal relay to bridge the two severed ends of an injured spinal cord.

Intercropping with sunflower and inoculation with arbuscular mycorrhizal fungi promotes growth of garlic chive in metal-contaminated soil at a WEEE-recycling site.

Heavy metal (HM) pollution in agricultural soils due to the recycling of waste electrical and electronic equipment (WEEE) has become a serious concern, but most farmers cannot afford the economic losses of fallow land during remediation. Thus, it is imperative to produce low-HM crops while remediating the contaminated soils. A 17-week pot experiment was conducted to investigate the growth and HM (Cd, Cu, Pb, Cr, Zn, and Ni) acquisition of garlic chives (Allium tuberosum Rottl. ex Spreng.) intercropped with sunflower (Helianthus annuus L.) and inoculated with (I+M) or without (I-M) the arbuscular mycorrhizal (AM) fungus Funneliformis caledonium on a severely HM-contaminated soil that was collected from a WEEE-recycling site. Compared with the monoculture control, the I-M treatment significantly (P < 0.05) decreased Cd, Cu, Cr, Zn, and Ni concentrations in the shoots of chives through rhizosphere competition and HM (except Cr) transfer from the root to the shoot of chives, and increased the average shoot fresh weight (i.e., yield) of chives by 794% by alleviating HM toxicity. Compared with the I-M treatment, the I+M treatment significantly increased soil phosphatase activity as well as root mycorrhizal colonization of both sunflower and chives. The I+M treatment had no effect on the tissue P concentration of sunflower but elevated the average dry biomass (shoot plus root) and P acquisition level of sunflower by 179% and 121%, respectively. In addition, the I+M treatment significantly increased the P concentration in the root rather than in the shoot of chives and significantly increased the level of P acquisition by chives, increasing the average yield of chives by 229%. Simultaneously, the I+M treatment significantly increased the level of HM (except Cd) acquisition by sunflower, enhancing the rhizosphere competition by sunflower over chives, and further reducing the transfer of all six HMs from root to shoot in the chives, and inducing significant decreases in chive shoot HM concentrations compared with the monoculture control. Furthermore, the I+M treatment decreased the average total concentrations and increased the average DTPA-extractable concentrations of soil HMs. The results demonstrate the multifunctional role of AM fungi in the intercropping system for both vegetable production and phytoremediation on HM-contaminated soils.

A Multifaceted Mass Spectrometric Method to Probe Feeding Related Neuropeptide Changes in Callinectes sapidus and Carcinus maenas.

Food intake is regulated by various neuromodulators, including numerous neuropeptides. However, it remains elusive at the molecular and cellular level as to how these important chemicals regulate internal processes and which regions of the neuronal organs are responsible for regulating the behavior. Here we report a comparative neuropeptidomic analysis of the brain and pericardial organ (PO) in response to feeding in two well-studied crustacean physiology model organisms, Callinectes sapidus and Carcinus maenas, using mass spectrometry (MS) techniques. A multifaceted MS-based approach has been developed to obtain complementary information on the expression changes of a large array of neuropeptides in the brain and PO. The method employs stable isotope labeling of brain and PO extracts for relative MS quantitation, capillary electrophoresis (CE)-MS for fractionation and high-specificity analysis, and mass spectrometric imaging (MSI) for in-situ molecular mapping of peptides. A number of neuropeptides, including RFamides, B-type allatostatins (AST-B), RYamides, and orcokinins exhibit significant changes in abundance after feeding in this investigation. Peptides from the AST-B family found in PO tissue were shown to have both altered expression and localization changes after feeding, indicating that they may be a class of vital neuropeptide regulators involved in feeding behavior. Graphical Abstract ᅟ.

The safety and pharmacokinetics of high dose intravenous ascorbic acid synergy with modulated electrohyperthermia in Chinese patients with stage III-IV non-small cell lung cancer.

Ascorbic acid (AA) infusion and modulated electrohyperthermia (mEHT) are widely used by integrative cancer practitioners for many years. However, there are no safety and pharmacokinetics data in Chinese cancer patients. We carried out a clinical trial to evaluate the safety and pharmacokinetics of those methods in patients with stage III-IV non-small cell lung cancer (NSCLC). Blood ascorbic acid in the fasting state was obtained from 35 NSCLC patients; selecting from them 15 patients with stage III-IV entered the phase I study. They were randomized allocated into 3 groups, and received doses 1.0, 1.2, 1.5g/kg AA infusions. Participants in the first group received intravenous AA (IVAA) when mEHT was finished, in the second group IVAA was administered simultaneously with mEHT and in the third group IVAA was applied first, and followed with mEHT. Pharmacokinetic profiles were obtained when they received solely IVAA and when IVAA in combination with mEHT. The process was applied 3 times a week (every other day, weekend days off) for 4weeks. We found that fasting plasma AA levels were significantly correlated with stage of the disease. Peak concentration of AA was significantly higher in the simultaneous treatments than in other combinations with mEHT or in solely IVAA-managed groups. IVAA synergy with simultaneous mEHT is safe and the concomitant application significantly increases the plasma AA level for NSCLC patients.

Tail Nerve Electrical Stimulation and Electro-Acupuncture Can Protect Spinal Motor Neurons and Alleviate Muscle Atrophy after Spinal Cord Transection in Rats.

Spinal cord injury (SCI) often results in death of spinal neurons and atrophy of muscles which they govern. Thus, following SCI, reorganizing the lumbar spinal sensorimotor pathways is crucial to alleviate muscle atrophy. Tail nerve electrical stimulation (TANES) has been shown to activate the central pattern generator (CPG) and improve the locomotion recovery of spinal contused rats. Electroacupuncture (EA) is a traditional Chinese medical practice which has been proven to have a neural protective effect. Here, we examined the effects of TANES and EA on lumbar motor neurons and hindlimb muscle in spinal transected rats, respectively. From the third day postsurgery, rats in the TANES group were treated 5 times a week and those in the EA group were treated once every other day. Four weeks later, both TANES and EA showed a significant impact in promoting survival of lumbar motor neurons and expression of choline acetyltransferase (ChAT) and ameliorating atrophy of hindlimb muscle after SCI. Meanwhile, the expression of neurotrophin-3 (NT-3) in the same spinal cord segment was significantly increased. These findings suggest that TANES and EA can augment the expression of NT-3 in the lumbar spinal cord that appears to protect the motor neurons as well as alleviate muscle atrophy.

Icariin attenuates titanium-particle inhibition of bone formation by activating the Wnt/ß-catenin signaling pathway in vivo and in vitro.

Wear-debris-induced periprosthetic osteolysis (PIO) is a common clinical condition following total joint arthroplasty, which can cause implant instability and failure. The host response to wear debris promotes bone resorption and impairs bone formation. We previously demonstrated that icariin suppressed wear-debris-induced osteoclastogenesis and attenuated particle-induced osteolysis in vivo. Whether icariin promotes bone formation in a wear-debris-induced osteolytic site remains unclear. Here, we demonstrated that icariin significantly attenuated titanium-particle inhibition of osteogenic differentiation of mesenchymal stem cells (MSCs). Additionally, icariin increased bone mass and decreased bone loss in titanium-particle-induced osteolytic sites. Mechanistically, icariin inhibited decreased ß-catenin stability induced by titanium particles in vivo and in vitro. To confirm icariin mediated its bone-protective effects via the Wnt/ß-catenin signaling pathway, we demonstrated that ICG-001, a selective Wnt/ß-catenin inhibitor, attenuated the effects of icariin on MSC mineralization in vitro and bone formation in vivo. Therefore, icariin could induce osteogenic differentiation of MSCs and promote new bone formation at a titanium-particle-induced osteolytic site via activation of the Wnt/ß-catenin signaling pathway. These results further support the protective effects of icariin on particle-induced bone loss and provide novel mechanistic insights into the recognized bone-anabolic effects of icariin and an evidence-based rationale for its use in PIO treatment.

Renoprotective effects of berberine as adjuvant therapy for hypertensive patients with type 2 diabetes mellitus: Evaluation via biochemical markers and color Doppler ultrasonography.

Diabetes and hypertension are complex and serious diseases that may ultimately lead to renal complications. Adequate control of blood glucose and blood pressure contributes to decreased renal risks, but may not be sufficient for certain patients. The current study was undertaken to investigate the renoprotective effects of berberine as an adjuvant therapy to standard hypotensive and hypoglycemic treatment in hypertensive patients with type 2 diabetes mellitus (T2DM). In this 2-year clinical study, 69 hypertensive patients with T2DM, whose blood pressure and fasting plasma glucose (FPG) were adequately controlled by hypotensive and oral hypoglycemic agents prior to the study, were enrolled and randomly assigned into control (33 cases) and add-on (36 cases) groups. Berberine was orally administrated to the patients in the add-on group concomitantly with standard hypotensive and hypoglycemic treatment. Baseline characteristics, including the levels of FPG, glycated hemoglobin, systolic blood pressure, diastolic blood pressure, serum creatinine, urinary albumin-to-creatine ratio (UACR), urinary osteopontin and kidney injury molecule-1 (KIM-1) were determined. Furthermore, the oxidative stress markers malondialdehyde, urinary 8-hydroxy-2'-deoxyguanosine, superoxide dismutase, glutathione peroxidase and total-antioxidant capacity, and the inflammatory parameters vascular adhesion molecule-1, C-reactive protein and high molecular weight-adiponectin were evaluated. In addition, ultrasonographic parameters, including peak systolic velocity, end diastolic velocity and renal arterial resistance index were determined. After treatment, it was observed that the control and add-on treatments were able to adequately control blood pressure and blood glucose. Patients in the add-on group exhibited significant reductions in renal damage biochemical markers (UACR, urinary osteopontin and KIM-1) and improved renal hemodynamics, in addition to reduced inflammation and oxidative stress. The present results suggest that berberine is beneficial for hypertensive patients with T2DM as add-on therapy to standard hypotensive and hypoglycemic agents.

Arbuscular mycorrhizal fungal diversity, root colonization, and soil alkaline phosphatase activity in response to maize-wheat rotation and no-tillage in North China.

Monitoring the effects of no-tillage (NT) in comparison with conventional tillage (CT) on soil microbes could improve our understanding of soil biochemical processes and thus help us to develop sound management strategies. The objective of this study was to compare the species composition and ecological function of soil arbuscular mycorrhizal (AM) fungi during the growth and rotation of crops under NT and CT. From late June 2009 to early June 2010, 32 topsoil (0-15 cm) samples from four individual plots per treatment (CT and NT) were collected at both the jointing and maturation stages of maize (Zea mays L.) and wheat (Triticum aestivum L.) from a long-term experimental field that was established in an Aquic Inceptisol in North China in June 2006. The AM fungal spores were isolated and identified and then used to calculate species diversity indices, including the Shannon- Wiener index (H'), Evenness (E), and Simpson's index (D). The root mycorrhizal colonization and soil alkaline phosphatase activity were also determined. A total of 34 species of AM fungi within nine genera were recorded. Compared with NT, CT negatively affected the soil AM fungal community at the maize sowing stage, leading to decreases in the average diversity indices (from 2.12, 0.79, and 0.82 to 1.79, 0.72, and 0.74 for H', E, and D, respectively), root mycorrhizal colonization (from 28% to 20%), soil alkaline phosphatase activity (from 0.24 to 0.19 mg/g/24 h) and available phosphorus concentration (from 17.4 to 10.5 mg/kg) at the maize jointing stage. However, reductions in diversity indices of H', E, and D were restored to 2.20, 0.81, and 0.84, respectively, at the maize maturation stage. CT should affect the community again at the wheat sowing stage; however, a similar restoration in the species diversity of AM fungi was completed before the wheat jointing stage, and the highest Jaccard index (0.800) for similarity in the species composition of soil AM fungi between CT and NT was recorded at the wheat maturation stage. Our results also demonstrated that NT resulted in the positive protection of the community structure of AM fungi and played an important role in maintaining their functionality especially for maize seedlings.

Molecular properties and immune defense of two ferritin subunits from freshwater pearl mussel, Hyriopsis schlegelii.

Ferritin is a conserved iron-binding protein involved in cellular iron metabolism and host defense. In the present study, two distinct cDNAs for ferritins in the freshwater pearl mussel Hyriopsis schlegelii were identified (designated as HsFer-1 and HsFer-2) by SMART RACE approach and expressed sequence tag (EST) analysis. The full-length cDNAs of HsFer-1 and HsFer-2 were of 760 and 877 bp, respectively. Both of the two cDNAs contained an open reading frame (ORF) of 522 bp encoding for 174 amino acid residues. Sequence characterization and homology alignment indicated that HsFer-1 and HsFer-2 had higher similarity to H-type subunit of vertebrate ferritins than L-type subunit. Analysis of the HsFer-1 and HsFer-2 untranslated regions (UTR) showed that both of them had an iron response element (IRE) in the 5'-UTR, which was considered to be the binding site for iron regulatory protein (IRP). Quantitative real-time PCR (qPCR) assays were employed to examine the mRNA expression profiles. Under normal physiological conditions, the expression level of both HsFer-1 and HsFer-2 mRNA were the highest in hepatopancreas, moderate in gonad, axe foot, intestine, kidney, heart, gill, adductor muscle and mantle, the lowest in hemocytes. After stimulation with bacteria Aeromonas hydrophila, HsFer-1 mRNA experienced a different degree of increase in the tissues of hepatopancreas, gonad and hemocytes, the peak level was 2.47-fold, 9.59-fold and 1.37-fold, respectively. Comparatively, HsFer-2 showed up-regulation in gonad but down-regulation in hepatopancreas and hemocytes. Varying expression patterns indicate that two types of ferritins in H. schlegelii might play different roles in response to bacterial challenge. Further bacteriostatic analysis showed that both the purified recombinant ferritins inhibited the growth of A. hydrophila to a certain degree. Collectively, our results suggest that HsFer-1 and HsFer-2 are likely to be functional proteins involved in immune defense against bacterial infection.

Mass spectral charting of neuropeptidomic expression in the stomatogastric ganglion at multiple developmental stages of the lobster Homarus americanus.

The stomatogastric nervous system (STNS) of the American lobster Homarus americanus serves as a useful model for studies of neuromodulatory substances such as peptides and their roles in the generation of rhythmic behaviors. As a central component of the STNS, the stomatogastric ganglion (STG) is rich in neuropeptides and contains well-defined networks of neurons, serving as an excellent model system to study the effect of neuropeptides on the maturation of neural circuits. Here, we utilize multiple mass spectrometry (MS)-based techniques to study the neuropeptide content and abundance in the STG tissue as related to the developmental stage of the animal. Capillary electrophoresis (CE)-MS was employed to unambiguously identify low abundance neuropeptide complements, which were not fully addressed using previous methods. In total, 35 neuropeptides from 7 different families were detected in the tissue samples. Notably, 10 neuropeptides have been reported for the first time in this study. In addition, we utilized a relative quantitation method to compare neuropeptidomic expression at different developmental stages and observed sequential appearance of several neuropeptides. Multiple isoforms within the same peptide family tend to show similar trends of changes in relative abundance during development. We also determined that the relative abundances of tachykinin peptides increase as the lobster grows, suggesting that the maturation of circuit output may be influenced by the change of neuromodulatory input into the STG. Collectively, this study expands our knowledge about neuropeptides in the crustacean STNS and provides useful information about neuropeptide expression in the maturation process.

[Effect on the motor function of stroke patients by combination of needling at Back-shu point and trunk exercise].

OBJECTIVE: To study the effect on the motor function of stroke patients by combination of needling at Back-shu point and trunk exercise. METHODS: Ninety stroke hemiplegic patients were randomly assigned to the conventional treatment group (as the convention group), the Back-shu point needling group, and the combination of Back-shu point needling and the trunk exercise group, 30 patients in each group. They were treated with the conventional treatment, needling at Back-shu point, and the combination of needling at Back-shu point and trunk exercise. The Fugl-Meyer score (FMA) and modified Barthel index (MBI) score were assessed before treatment and two months after treatment. RESULTS: The three rehabilitation treatment methods were all effective in improving the motor function of stroke hemiplegic patients (P<0.05). The effects in the Back-shu point needling group and the combination of Back-shu point needling and the trunk exercise group were respectively superior to that in the conventional treatment group (P<0.05). The effect in the combination of Back-shu point needling and the trunk exercise group was superior to that in the Back-shu point needling group (P<0.05). CONCLUSION: The combination of Back-shu point needling and the trunk exercise could improve the motor function of stroke hemiplegic patients, and its effect was better than needling at Back-shu point alone.

Arbuscular mycorrhizal fungus enhances P acquisition of wheat (Triticum aestivum L.) in a sandy loam soil with long-term inorganic fertilization regime.

The P efficiency, crop yield, and response of wheat to arbuscular mycorrhizal fungus (AMF) Glomus caledonium were tested in an experimental field with long-term (19 years) fertilizer management. The experiment included five fertilizer treatments: organic amendment (OA), half organic amendment plus half mineral fertilizer (1/2 OM), mineral fertilizer NPK, mineral fertilizer NK, and the control (without fertilization). AMF inoculation responsiveness (MIR) of wheat plants at acquiring P were estimated by comparing plants grown in unsterilized soil inoculated with G. caledonium and in untreated soil containing indigenous AMF. Without AMF inoculation, higher crop yields but lower colonization rates were observed in the NPK and two OA-inputted treatments, and NPK had significantly (P < 0.05) lower impacts on organic C and available P in soils and thereby P acquisition of wheat plants compared with OA and 1/2 OM. G. caledonium inoculation significantly (P < 0.05) increased colonization rates with the NPK and two P-deficient treatments but significantly (P < 0.05) increased vegetative biomass, crop yield, and P acquisition of wheat as well as soil alkaline phosphatase (ALP) activity, only with the NPK treatment. This gave an MIR of ca. 45% on total P acquisition of wheat plants. There were no other remarkable MIRs. It suggested that the MIR is determined by soil available P status, and rational combination of AMF with chemical NPK fertilizer can compensate for organic amendments by improving P-acquisition efficiency in arable soils.

[Efficacy of integrative respiratory rehabilitation training in exercise ability and quality of life of patients with chronic obstructive pulmonary disease in stable phase: a randomized controlled trial].

BACKGROUND: Chronic obstructive pulmonary disease (COPD) is an important clinical disease, and its global prevalence and mortality rates are high. It is meaningful to investigate the efficacy of integrative respiratory rehabilitation training in quality of life and respiratory physiology of COPD patients in stable phase. OBJECTIVE: To observe the efficacy of integrative respiratory rehabilitation training in exercise ability and quality of life of COPD patients in stable phase. DESIGN, SETTING, PARTICIPANTS AND INTERVENTIONS: Eighty outpatients and inpatients with COPD from Department of Respiratory Medicine, Taihe Hospital, Yunyang Medical College were randomly divided into 4 groups, with 20 patients in each group. The patients in group A only received drug therapy, the patients in group B received traditional qigong training, the patients in group C received modern rehabilitation training, and the patients in group D received integrative respiratory rehabilitation training. MAIN OUTCOME MEASURES: Chronic respiratory questionnaire (CRQ), 6-minute walking distance and Borg score in each group were examined before and after one-, three-, and six-month and one-year treatment. RESULTS: The 6-minute walking distance, Borg score and CRQ score in group A had no significant changes after treatment (P>0.05). After one-month treatment, there were no significant differences in 6-minute walking distance and Borg score in groups B, C and D as compared with those before treatment (P<0.05). After three-month treatment, 6-minute walking distance and Borg score were improved in groups B, C and D (P<0.05). After six-month and one-year treatment, 6-minute walking distance, Borg score and CRQ score in groups B, C and D were improved as compared with those before treatment (P<0.05), and there were significant differences between group D and any of groups A, B and C (P<0.05). CONCLUSION: Modern rehabilitation training, traditional qigong training and integrative respiratory rehabilitation training programs all can improve the quality of life and exercise ability of COPD patients, and integrative respiratory rehabilitation training program is better than modern rehabilitation training and traditional qigong training programs. The efficacy of respiratory rehabilitation training is time-dependent, and need long-time adherence to the therapy.

[Effect of electroacupuncture of different acupoints on the excitability of detrusor muscle and the expression of BDNF and TrkB in the spinal cord of rats with urinary retention due to spinal cord injury].

OBJECTIVE: To observe the effect of electroacupuncture (EA) of different acupoints on the expression of brain-derived neurotrophic factor (BDNF) and tropomyosine receptor kinase B (TrkB) in the spinal cord and the excitability of detrusor muscle of the uninary bladder in rats with urinary retention owing to spinal cord injury (SCI). METHODS: A total of 100 female SD rats were randomly divided into normal, sham-operation (sham), model, EA-Guanyuan (CV 4), and EA-Shuidao (ST 28) groups, with 20 cases in each. SCI induced urinary retention model was established by using weight dropping method. EA (1 mA, 2 Hz/15 Hz) was applied to "Guanyuan" (CV 4) and "Shuidao" (ST 28) respectively for 20 min, once a day for 10 days. The excitability (tone, contraction frequency) of the detrusor muscle of the bladder was detected in vitro by using electrophysiological method, and the expression of BDNF and TrkB in spinal cord was determined by immunohistochemistry staining. RESULTS: In comparison with normal control and sham groups, the tension and the contraction frequency of detrusor muscle in model group lowered significantly (P<0.05), while compared with model group, both the tension and contraction frequency of detrusor muscle increased pronouncedly in EA-CV 4 and EA-ST 28 groups (P<0.05), and the effect of EA-CV 4 was apparently superior to that of EA-ST 28 (P<0.05). In comparison with normal and sham groups, the BDNF and TrkB immunoreaction positive cells in the spinal cord were significantly more in model group (P<0.05). Compared with model group, those of EA-CV 4 and EA-ST 28 groups were obviously further increased (P<0.05), and the effect of EA-CV 4 group was markedly superior to that of EA-ST 28 group (P<0.05). CONCLUSION: EA of CV 4 and ST 28 can raise the excitability of the smooth muscle of the uninary bladder in rats with SCI-induced urinary retention, which may be related to its effects in upregulating the expression of BDNF and TrkB in the spinal cord. The effects of EA of CV 4 were evidently superior to those of EA of ST 28.

[Effects of earthworm (Eisennia foetida ) on As, P uptake by maize and As, P fractional transformation in the rhizosphere].

A pot experiment was conducted to examine the roles of earthworm in As uptake from original As-polluted soil by maize (Zea mays L.), and their effects on As, P fractions in the rhizosphere. The As-polluted soils with three As levels were collected from the arable soil near As mine. The plants were harvested after 10 weeks of growth. Dry weight (DW) and P, As concentrations of plants, as well as As and P fractions of the rhizospheric soil, were determined. The results showed that inoculated earthworm or appended rice straw increased maximal 149%, 222% DW of root and shoot, respectively. At the medium and high soil As levels, root As concentration in the soil treated by earthworm and rice straw was highest among all treatments, and earthworm increased more As concentration of shoot than rice straw did. In different soil As levels, root P concentration in the soil treated by earthworm was highest, and shoot P by rice straw. Ca-P affected maize absorbing As at the low soil As level(r = 0.981), and maize absorbing Al-P was restrained by As involved in well-crystallized hydrous oxides of Fe and Al at the medium (r = 0.953)and high (r = 0.997)soil As levels. The concentration of non-specially absorbed As and As combined with Fe or Al and of O-P increased at the soil inoculated earthworm or/and appended rice straw at the same time. These results indicated that earthworm was more valuable for plant developing than rice straw was.