Potential low toxic alternative for Na-Cl cotransporter inhibition: A diuretic effect and mechanism study of Pyrrosia petiolosa.

BACKGROUND: Hydrochlorothiazide, a diuretic commonly used for the treatment of hypertension, is often associated with serious metabolic side effects. Pyrrosia petiolosa (Christ) Ching is a traditional Chinese medicine that possesses diuretic properties, without any obvious side effects. AIM: To evaluate the diuretic effect of P. petiolosa (Christ) Ching and to elucidate its underlying mechanism of action. METHODS: Extracts obtained from different polar components of P. petiolosa (Christ) Ching were analyzed for toxicity in a Kunming mouse model. The diuretic effects of the extracts were compared to that of hydrochlorothiazide in rats. In addition, compound isolation procedures, cell assays of Na-Cl cotransporter inhibition and rat diuretic test of monomeric compounds were conducted to identify the active ingredients in the extract. Subsequently, homology modeling and molecular docking were performed to explain the reason behind the diuretic activity observed. Finally, LC-MS analysis was used to elucidate the underlying mechanism of action of P. petiolosa (Christ) Ching. RESULTS: No toxicity was observed in mice administered P. petiolosa (Christ) Ching extracts. The ethyl acetate fraction showed the most significant diuretic effect. Similar results were obtained during the analysis for Na+ content in rat urine. Further separation of P. petiolosa (Christ) Ching components led to the isolation of methyl chlorogenate, 2',3'-dihydroxy propyl pentadecanoate, and ß-carotene. Results from cell assays showed that the Na-Cl cotransporter inhibitory activity of methyl chlorogenate was greater than that of hydrochlorothiazide. This result was again confirmed by the diuresis tests of monomeric compounds in rats. The molecular simulations explain the stronger interactions between the methyl chlorogenate and Na-Cl cotransporter. Of the compounds determined using LC-MS analysis, 185 were identified to be mostly organic acids. CONCLUSIONS: P. petiolosa possesses significant diuretic activities without any obvious toxicity, with least two possible mechanisms of action. Further study on this herb is warranted.

Effects of citric acid supplementation on growth performance, intestinal morphology and microbiota, and blood parameters of geese from 1 to 28 days of age.

This study was conducted to investigate the effects of citric acid (CA) supplementation on growth performance, intestinal morphology, intestinal microbiota, and blood parameters of geese from 1 to 28 d of age and evaluate the optimum additional level of citric acid. A total of 180 one-day-old male goslings were randomly allotted to 5 treatment groups of 36 birds with 6 replications. The control group was fed a basal diet, and the other groups were fed the basal diet supplemented with 0.25, 0.50, 1.00, and 2.00% of citric acid, respectively. The results showed that goslings fed the diet supplemented with 1.00% CA had higher final body weight (FBW) and average daily gain (ADG) than other groups (P < 0.05). The CA supplementation at 0.25 to 1.00% improved the morphology of duodenum or jejunum (P < 0.05). The jejunal content pH value was significantly reduced with the addition of CA compared with the control group (P < 0.05). As citric acid levels increased, the IgA concentrations in plasma increased and then decreased, and the goslings fed 1.00% CA supplementation had the highest IgA concentrations (P < 0.05). The supplementation of 1.00% and 2.00% CA in diet significantly reduced the malondialdehyde (MDA) concentration in plasma (P < 0.05). No significant difference was found on some indices related to liver function in plasma (P > 0.05), while creatinine significantly increased by the 2.00% CA supplementation (P < 0.05). Besides, the higher Coliform level in cecal content and worse intestinal morphology were observed when CA supplementation was up to 2.00%. Hence, the dietary CA supplementation (especially 1.00%) in goslings improves the growth performance, intestinal morphology, immunity and antioxidant, while excessive CA addition may cause negative effects. According to the quadratic polynomial model, the addition of CA in diet for obtaining maximum average daily feed intake (ADFI) should be 1.09% (10.9 g/kg diet) for goslings from 1 to 28 d of age.

Reducing postsurgical exudate in breast cancer patients by using San Huang decoction to ameliorate inflammatory status: a prospective clinical trial.

Background: Reducing inflammatory factors in wound exudate is a promising treatment approach for healing wounds in postsurgical breast cancer patients. Traditional Chinese Medicine (tcm) treatments have been shown to be beneficial and safe for optimal regulation of oxidative stress during the postoperative period. In the present clinical trial, we evaluated the effectiveness of a promising Chinese herbal formula, San Huang decoction [shd (Radix astragali, Radix et rhizoma rhei, and Rhizoma curcuma longa, 3:1:1; supplemental Table 1)], on wound inflammatory response after mastectomy. Methods: The study randomized 30 patients with breast cancer who fulfilled the inclusion and exclusion criteria to either a treatment (n = 15) or a control group (n = 15). Patients in the treatment group received liquid shd, taken twice daily with or without food. Treatment was given for 1 day before surgery and for 7 days postoperatively. Participants in the control group received a placebo on the same schedule as the treatment group. Outcomes measured in every subject included clinical tcm and wound inflammation symptom scores, daily and total amounts of drainage fluid, and levels of inflammatory factors in the exudate [tumour necrosis factor α (tnf-α), interleukins 6 (il-6), 8 (il-8), and 2R (il-2R), human C-reactive protein (crp)] at 2 hours and on days 1, 3, and 7 postoperatively. Results: The total amount of drainage fluid over 7 days was significantly lower in the treatment group (572.20 ± 93.95 mL) than in the control group (700.40 ± 107.38 mL). The tcm symptom score was also lower in treatment group (day 7: 1.87 ± 0.83 vs. 4.80 ± 3.61, p = 0.049), as was the inflammatory symptom score (day 7: 0.67 ± 0.72 vs. 3.67 ± 2.50, p = 0.001). Levels of tnf-α, il-6, il-8, il-2R, and crp in drainage fluid were significantly lower with shd treatment. Conclusions: Perioperative treatment with shd effectively lessened postoperative exudate and ameliorated inflammatory symptoms in patients who underwent surgery for breast cancer.

Simultaneous determination of organochlorine and pyrethriod pesticide residues in the Chinese patent medicines by gas chromatography-tandem mass spectrometry.

A simple, sensitive, reliable method was developed for the simultaneous determination of organochlorine and pyrethriod pesticide residues in Chinese patent medicines Six ingredient rehmannia pills and Xiaoyao pills. These pesticides were extracted by ethyl acetate. The extraction time and volume of ethyl acetate were optimized. Cleanup of extracts was performed with dispersive-solid phase extraction using graphitized carbon black as the sorbent. The determination of pesticides in the final extracts was carried out by gas chromatography-tandem mass spectrometry in multiple reaction monitoring mode (GC-MS/MS, MRM). The linearity of the calibration curves is good in matrix-matched standard and yields the coefficients of determination (R2) ≥0.99 for all of the target analytes. Under optimized conditions, the average recoveries (five replicates) for most pesticides range from 75.5% to 114.6%, and RSDs are less than 10.0%. The LODs of 18 pesticides in Six ingredient rehmannia pill and Xiaoyao pills are in the range of 0.01-8.82 µg kg-1. The developed method meets the requirements of pesticide residue analysis and could be effectively used for routine analysis of the organochlorine and pyrethriod pesticide residues in Six ingredient rehmannia pills and Xiaoyao pills.

Effects of excess DL- and L-methionine on growth performance of starter Pekin ducks.

A dose-response experiment was conducted to investigate the effects of excess DL-methionine (DLM) and L-methionine (LM) on growth response of starter Pekin ducks from 7 to 21 days of age. A total of 462 seven-day-old male Pekin ducklings were allotted to 77 wire-floor pens with 6 birds per pen. There were 11 treatments including a methionine-adequate control diet (containing 0.49% methionine) and control diets supplemented with 5 levels of crystal DLM or LM (0.25, 0.50, 0.75, 1.0, and 1.25%) based on equal product weight. At 21 d of age, weight gain, feed intake, and feed/gain of ducks from each pen were measured. In our study, the weight gain and feed intake did not change markedly and kept a plateau when supplemental DLM or LM was below 0.50%, but the weight gain and feed intake decreased further as supplemental DLM or LM increased from 0.50 to 1.25%. When the maximum safe level (MSL) of supplemental methionine sources and methionine content of the control diets were combined, the total tolerable upper limits of methionine for weight gain and feed intake were 0.91 and 0.85% when DLM was used, respectively, and the corresponding values were 0.89 and 0.84% when LM was used, respectively. On the other hand, both excess DLM and LM reduced weight gain and feed intake (P < 0.05) but there were no significant differences (P > 0.05) in weight gain and feed intake between the ducks fed DLM- and LM-supplemented diets. Furthermore, linear slope-ratio assay was also used to evaluate the relative toxicity of excess DLM and LM. According to this assay, the efficiencies of DLM relative to LM for depression of weight gain and feed intake were 97 and 95%, respectively. In summary, excess DL- and L-methionine were toxic for starter Pekin ducks and both methionine sources were equally growth depressing.

Clinical outcomes of extended versus intermittent administration of piperacillin/tazobactam for the treatment of hospital-acquired pneumonia: a randomized controlled trial.

The purpose of this study was to assess the pharmacokinetic (PK) characteristics, clinical efficiency, and pharmacoeconomic parameters of piperacillin/tazobactam administered by extended infusion (EI) or intermittent infusion (II) in the treatment of hospital-acquired pneumonia (HAP) in critically ill patients with low illness severity in China. Fifty patients completed the study, with 25 patients receiving 4/0.5 g piperacillin/tazobactam over 30 min as the II group and 25 patients receiving 4/0.5 g piperacillin/tazobactam over 3 h every 6 h as the EI group. Drug assay was performed using high-performance liquid chromatography (HPLC). The percentage of the dosing interval for which the free piperacillin concentration (%fT) exceeds the minimum inhibitory concentration (MIC) was calculated. The patients' therapy cost, clinical efficiency, and adverse effects were also recorded. %fT>MIC was about 100, 98.73, and 93.04 % in the EI arm versus 81.48, 53.29, and 42.15 % in the II arm, respectively, when the microorganism responsible for HAP had an MIC of 4, 8, and 16 mg/L. The therapy cost in the EI group was lower than that of the II group ($1351.72 ± 120.39 vs. $1782.04 ± 164.51, p = 0.001). However, the clinical success rate, clinical failure rate, and drug-related adverse events did not significantly differ between groups. EI treatment with piperacillin/tazobactam was a cost-effective approach to the management of HAP, being equally clinically effective to conventional II.

Analysis of genetic diversity of Brassica rapa var. chinensis using ISSR markers and development of SCAR marker specific for Fragrant Bok Choy, a product of geographic indication.

Non-heading Chinese cabbage [Brassica rapa var. chinensis (Linnaeus) Kitamura] is a popular vegetable and is also used as a medicinal plant in traditional Chinese medicine. Fragrant Bok Choy is a unique accession of non-heading Chinese cabbage and a product of geographic indication certified by the Ministry of Agriculture of China, which is noted for its rich aromatic flavor. However, transitional and overlapping morphological traits can make it difficult to distinguish this accession from other non-heading Chinese cabbages. This study aimed to develop a molecular method for efficient identification of Fragrant Bok Choy. Genetic diversity analysis, based on inter-simple sequence repeat molecular markers, was conducted for 11 non-heading Chinese cabbage accessions grown in the Yangtze River Delta region. Genetic similarity coefficients between the 11 accessions ranged from 0.5455 to 0.8961, and the genetic distance ranged from 0.0755 to 0.4475. Cluster analysis divided the 11 accessions into two major groups. The primer ISSR-840 amplified a fragment specific for Fragrant Bok Choy. A pair of specific sequence-characterized amplified region (SCAR) primers based on this fragment amplified a target band in Fragrant Bok Choy individuals, but no band was detected in individuals of other accessions. In conclusion, this study has developed an efficient strategy for authentication of Fragrant Bok Choy. The SCAR marker described here will facilitate the conservation and utilization of this unique non-heading Chinese cabbage germplasm resource.

Transcriptomic analysis of Camellia ptilophylla and identification of genes associated with flavonoid and caffeine biosynthesis.

Camellia ptilophylla, or cocoa tea, is naturally decaffeinated and its predominant catechins and purine alkaloids are trans-catechins and theobromine Regular tea [Camellia sinensis (L.) O. Ktze.] is evolutionarily close to cocoa tea and produces cis-catechins and caffeine. Here, the transcriptome of C. ptilophylla was sequenced using the 101-bp paired-end technique. The quality of the raw data was assessed to yield 70,227,953 cleaned reads totaling 7.09 Gbp, which were assembled de novo into 56,695 unique transcripts and then clustered into 44,749 unigenes. In catechin biosynthesis, leucoanthocyanidin reductase (LAR) catalyzes the transition of leucoanthocyanidin to trans-catechins, while anthocyanidin synthase (ANS) and anthocyanidin reductase (ANR) catalyze cis-catechin production. Our data demonstrate that two LAR genes (CpLAR1 and CpLAR2) by C. ptilophylla may be advantageous due to the combined effects of this quantitative trait, permitting increased leucoanthocyanidin consumption for the synthesis of trans-catechins. In contrast, the only ANS gene observed in C. sinensis (CsANS) shared high identity (99.2%) to one homolog from C. ptilophylla (CpANS1), but lower identity (~80%) to another (CpANS2). We hypothesized that the diverged CpANS2 might have lost its ability to synthesize cis-catechins. C. ptilophylla and C. sinensis each contain two copies of ANR, which share high identity and may share the same function. Transcriptomic sequencing captured two N-methyl nucleosidase genes named NMT1 and NMT2. NMT2 was highly identical to three orthologous genes TCS2, PCS2, and ICS2, which did not undergo methylation in vitro; in contrast, NMT1 was less identical to TCS, PCS and ICS, indicating that NMT1 may undergo neofunctionalization.

Alveolar bone regeneration potential of a traditional Chinese medicine, Bu-Shen-Gu-Chi-Wan, in experimental periodontitis.

BACKGROUND AND OBJECTIVE: The aim of this study was to investigate the effect of a traditional Chinese medicine, Bu-Shen-Gu-Chi-Wan, on experimental periodontitis and bone regeneration in rat. MATERIAL AND METHODS: Sixty female Sprague-Dawley rats were divided equally into three groups: a healthy control group (Group N); a periodontitis group (Group P); and the Bu-Shen-Gu-Chi-Wan treatment group (Group T). A 0.2-mm wire was placed around the maxillary first molar and Porphyromonas gingivalis was injected into the gingival sulcus. Rats in different groups were administered 0.9% normal saline or Bu-Shen-Gu-Chi-Wan solution (4 g/kg of body weight, for three alternate days), and the animals were killed after 4 wk. Morphological analysis of alveolar bone rebuilding was performed using micro-computed tomography (micro-CT) and stereomicroscopy, and the variation of inflammation in the periodontium was determined histologically. The serum levels of the proinflammatory cytokines interleukin-1beta (IL-1ß) and tumor necrosis factor-alpha (TNF-α) and of the bone-turnover biomarkers pyridinoline cross-linked carboxyterminal telopeptide of type I collagen (ICTP) and osteocalcin (OC) were determined using radioimmunoassays. RESULTS: After treatment with Bu-Shen-Gu-Chi-Wan, there were significant decreases in the levels of IL-1ß, TNF-α, ICTP and OC and decreased inflammatory infiltration in the periodontal tissues of Group T. significant changes in alveolar bone volume and density were detected by micro-CT, but stereomicroscopy did not detect a significant improvement of alveolar bone height. CONCLUSION: The data of the present study suggest that the traditional Chinese medicine, Bu-Shen-Gu-Chi-Wan, has anti-inflammatory function in experimental periodontitis and may simultaneously improve alveolar bone remodeling.

Epigenetic synergies between biotin and folate in the regulation of pro-inflammatory cytokines and repeats.

The protein biotin ligase, holocarboxylase synthetase (HLCS), is a chromatin protein that interacts physically with the DNA methyltransferase DNMT1, the methylated cytosine-binding protein MeCP2 and the histone H3 K9-methyltransferase EHMT1, all of which participate in folate-dependent gene repression. Here we tested the hypothesis that biotin and folate synergize in the repression of pro-inflammatory cytokines and long-terminal repeats (LTRs), mediated by interactions between HLCS and other chromatin proteins. Biotin and folate supplementation could compensate for each other's deficiency in the repression of LTRs in Jurkat and U937 cells. For example, when biotin-deficient Jurkat cells were supplemented with folate, the expression of LTRs decreased by >70%. Epigenetic synergies were more complex in the regulation of cytokines compared with LTRs. For example, the abundance of TNF-α was 100% greater in folate- and biotin-supplemented U937 cells compared with biotin-deficient and folate-supplemented cells. The NF-κB inhibitor curcumin abrogated the effects of folate and biotin in cytokine regulation, suggesting that transcription factor signalling adds an extra layer of complexity to the regulation of cytokine genes by epigenetic phenomena. We conclude that biotin and folate synergize in the repression of LTRs and that these interactions are probably mediated by HLCS-dependent epigenetic mechanisms. In contrast, synergies between biotin and folate in the regulation of cytokines need to be interpreted in the context of transcription factor signalling.

Highly selective microbial transformation of major ginsenoside Rb1 to gypenoside LXXV by Esteya vermicola CNU120806.

AIMS: This study examined the biotransformation pathway of ginsenoside Rb(1) by the fungus Esteya vermicola CNU 120806. METHODS AND RESULTS: Ginsenosides Rb(1) and Rd were extracted from the root of Panax ginseng. Liquid fermentation and purified enzyme hydrolysis were employed to investigate the biotransformation of ginsenoside Rb(1) . The metabolites were identified and confirmed using NMR analysis as gypenoside XVII and gypenoside LXXV. A mole yield of 95·4% gypenoside LXXV was obtained by enzymatic conversion (pH 5·0, temperature 50°C). Ginsenoside Rd was used to verify the transformation pathway under the same reaction condition. The product Compound K (mole yield 49·6%) proved a consecutive hydrolyses occurred at the C-3 position of ginsenoside Rb(1) . CONCLUSIONS: Strain CNU 120806 showed a high degree of specific ß-glucosidase activity to convert ginsenosides Rb(1) and Rd to gypenoside LXXV and Compound K, respectively. The maximal activity of the purified glucosidase for ginsenosides transformation occurred at 50°C and pH 5·0. Compared with its activity against pNPG (100%), the ß-glucosidase exhibited quite lower level of activity against other aryl-glycosides. Enzymatic hydrolysate, gypenoside LXXV and Compound K were produced by consecutive hydrolyses of the terminal and inner glucopyranosyl moieties at the C-3 carbon of ginsenoside Rb(1) and Rd, giving the pathway: ginsenoside Rb(1) → gypenoside XVII → gypenoside LXXV; ginsenoside Rd→F(2) →Compound K, but did not hydrolyse the 20-C, ß-(1-6)-glucoside of ginsenoside Rb(1) and Rd. SIGNIFICANCE AND IMPACT OF THE STUDY: The results showed an important practical application on the preparation of gypenoside LXXV. Additionally, this study for the first time provided a high efficient preparation method for gypenoside LXXV without further conversion, which also gives rise to a potential commercial enzyme application.

Transcutaneous electrical nerve stimulation on acupoints reduces fentanyl requirement for postoperative pain relief after total hip arthroplasty in elderly patients.

BACKGROUND: Transcutaneous electrical nerve stimulation (TENS) is regarded as an effective treatment for various types of pain. However, no randomized controlled trial has investigated TENS on acupoints for postoperative analgesia in elderly patients. This study aim to investigate whether TENS on acupoints has any favorable effect on complementary analgesia after total hip arthroplasty (THA) for elderly patients compared with a sham control treatment. METHODS: Sixty-eight elderly patients requiring THA surgery were enrolled and randomly allocated to one of two groups. Group Acu received true TENS on acupoints (bilateral P6, L14; ST36, GB31 ipsilateral to the surgery site) and Group Sham received sham treatment. All patients received patient-controlled analgesia for two days postoperatively. Analgesia was assessed by postoperative fentanyl requirement and pain intensity using a visual analogue scale (VAS-10 cm). The incidence of analgesia-related side effects, optional medication use and effects of patients' blinding were recorded. RESULTS: Fentanyl consumption in Group Acu was lower than that in Group Sham at 24 h (mean ± SD; 360±117 vs. 572±132 µg; P<0.001) and 48 h (712±184 vs. 1022±197 µg; P<0.001) after surgery. Postoperative pain intensity measured by VAS was similar in both groups. The incidence of opioid-related side effects and rescue medication for postoperative analgesia was significantly higher in Group Sham than in Group Acu. Differences between the groups regarding the effects of patients' blinding were not significant. CONCLUSION: TENS on specific acupoints is an effective and complementary approach to reduce postoperative analgesic requirement in elderly patients after THA.

The effects of electromechanical wrist robot assistive system with neuromuscular electrical stimulation for stroke rehabilitation.

An electromyography (EMG)-driven electromechanical robot system integrated with neuromuscular electrical stimulation (NMES) was developed for wrist training after stroke. The performance of the system in assisting wrist flexion/extension tracking was evaluated on five chronic stroke subjects, when the system provided five different schemes with or without NMES and robot assistance. The tracking performances were measured by range of motion (ROM) of the wrist and root mean squared error (RMSE). The performance is better when both NMES and robot assisted in the tracking than those with either NMES or robot only (P<0.05). The muscle co-contractions in the upper limb measured by EMG were reduced when NMES provided assistance (P<0.05). All subjects also attended a 20-session wrist training for evaluating the training effects (3-5 times/week). The results showed improvements on the voluntary motor functions in the hand, wrist and elbow functions after the training, as indicated by the clinical scores of Fugl-Meyer Assessment, Action Research Arm Test, Wolf Motor Function Test; and also showed reduced spasticity in the wrist and the elbow as measured by the Modified Ashworth Score of each subject. After the training, the co-contractions were reduced between the flexor carpi radialis and extensor carpi radialis, and between the biceps brachii and triceps brachii. Assistance from the robot helped improve the movement accuracy; and the NMES helped increase the muscle activation for the wrist joint and suppress the excessive muscular activities from the elbow joint. The NMES-robot assisted wrist training could improve the hand, wrist, and elbow functions.

First Report of Subanguina moxae Infecting Mugwort in Yunnan, China.

Mugwort is a perennial in the Compositae family distributed throughout Asia and Europe. The leaves are reported to have various pharmaceutical properties, e.g., antibacterial, antiviral, antitussive, and hemostatic properties, and have been used in traditional Chinese medicine for more than 2,000 years. In August 2011, a field of mugwort in Kunming, Yunnan Province, China, exhibited more than 90% incidence of whitish and rounded galls on the leaves. There were approximately 10 galls on each leaf, impacting the quality of the leaves for medicinal use. Parasitic nematodes were found upon dissection of the galls, then eggs, second-stage juveniles (J2), and mature males and females were observed. Through the morphologic observation of juveniles and female and male adults, the parasitic nematode was identified as Subanguina moxae (Yokoo and Choi, 1968) Brzeski, 1981 (3). Key morphological features are as follows: eggs (n = 20) measured 54.0 to 71.4 × 24.1 to 30.0 µm; J2 (n = 20) had the following characteristics: body length 689.3 to 873.2 µm (x = 775.5 µm); stylet length 8.2 to 9.8 µm (x = 8.8 µm); tail length 49.5 to 74.5 µm (x = 60.1 µm); a (total body length/maximum body width) ranged from 28.6 to 38.6 µm (x = 34.1 µm); and c (total body length/the length of the tail) ranged from 11.2 to 16.0 µm (x = 13.0 µm). Females (n = 20) had the following characteristics: body length 1,252.8 to 1,665.2 µm (x = 1,475.7 µm); stylet length 7.2 to 9.2 µm (x = 8.2 µm); V of 88.0 to 92.3 µm (x = 89.6 µm); a ranged from 17.6 to 24.5 µm (x = 21.3 µm); and c ranged from 20.2 to 28.9 µm (x = 22.8 µm). Males (n = 20) had the following characteristics: body length 994.2 to 1,453.6 µm (x = 1,253.2 µm); stylet length 7.5 to 9.9 µm (x= 9.1 µm); tail length 69.2 to 88.1 µm (x = 78.0 µm); spicule length 22.2 to 33.4 µm (x = 29.4 µm); gubernaculum length 10.4 to 14.2 µm (x = 12.2 µm); a ranged from 23.1 to 37.2 µm (x = 29.9 µm); and c ranged from 13.9 to 18.7 µm (x = 16.1 µm). Amplification of the rDNA-internal transcribed spacer (ITS) region and the D2/D3 (1) fragments of the 28S RNA with universal primers rDNA1/rDNA2 and D2A/D3B yielded PCR fragments of 934 bp and 754 bp, respectively. The ITS sequence (JN865234) and D2D3 sequence (JN885540) were submitted to GenBank. The ITS sequence (JN865234) exhibited 99.4% similarity with Mesoanguina moxae (AF396314) (synonym of S. moxae) (4). S. moxae has been identified from the common mugwort in Japan (2) and in China, was reported to infect wheat in Guizhou Province, but to our knowledge, this is the first report of this nematode affecting mugwort in Yunnan, China. References: (1) S. Amiri et al. Eur. J. Plant Pathol. 108:497, 2002. (2) K. Daigo et al. Bull. School Agric. Meiji University. 56:237, 2007. (3) M. R. Siddiqi. Tylenchida: Parasites of Plants and Insects. CABI Publishing, New York, 2000. (4) S. A. Subbotin et al. Mol. Phylogenet. Evol. 30:226, 2004.

Autophagy inhibition contributes to radiation sensitization of esophageal squamous carcinoma cells.

Radiotherapy is a useful component of treatment strategies for esophageal cancer. The role of autophagy in response to ionizing radiation was investigated in human esophageal squamous carcinoma cells. Cell viability and clonogenic survival assay were used to evaluate the radiosensitivity of autophagy inhibitor (3-MA) on esophageal squamous carcinoma cells. The percentage of apoptotic cells and cell cycle analysis were assessed by flow cytometry; DAPI staining was used to detect apoptotic cells. The expression of beclin-1 and LC3 was measured using a Western blot. The ultrastructural analysis was under the electron microscope. 6 Gy irradiation induced a massive accumulation of autophagosomes accompanied by strong upregulation of beclin-1 and LC3-II expression in TE-1 cells. Compared with radiation alone, 3-MA combined with radiation significantly decreased cell viability, as well as autophagic ratio, beclin-1, and LC3-II protein level. Inhibition of autophagy increased radiation-induced apoptosis and the percentage of G2/M-phase cells. Blockade of autophagy with 3-MA enhanced cytotoxicity of radiotherapy in human esophageal squamous carcinoma cells. It suggests that inhibition of autophagy could be used as adjuvant therapy to treat esophageal squamous cell carcinoma.

Effectiveness of functional electrical stimulation (FES)-robot assisted wrist training on persons after stroke.

A functional electrical stimulation (FES)-robot system controlled by subjects' motor intention was developed in our previous study. The effectiveness of the FES-robot on wrist training was investigated in this work. Five hemiplegic subjects with chronic stroke were recruited for an FES-robot assisted wrist training with 20 sessions. After the training, motor improvements were found in the wrist and fingers, represented by significant increase (P < 0.05) in clinical scores of the Fugl-Meyer Assessment (FMA), the Action Research Arm Test (ARAT), and the Modified Ashworth Score (MAS). Muscle coordination in the upper limb was also improved during the training as assessed by electromyography. The increased ARAT scores suggested improved upper limb motor functions, especially in the hand and fingers, compared to no improvement in previous study with only interactive robot-assisted wrist training without FES.

Cerebral magnetic resonance imaging of compressed air divers in diving accidents.

To investigate the characteristics of the cerebral magnetic resonance imaging (MRI) of compressed air divers in diving accidents, we conducted an observational case series study. MRI of brain were examined and analysed on seven cases compressed air divers complicated with cerebral arterial gas embolism CAGE. There were some characteristics of cerebral injury: (1) Multiple lesions; (2) larger size; (3) Susceptible to parietal and frontal lobe; (4) Both cortical grey matter and subcortical white matter can be affected; (5) Cerebellum is also the target of air embolism. The MRI of brain is an sensitive method for detecting cerebral lesions in compressed air divers in diving accidents. The MRI should be finished on divers in diving accidents within 5 days.

Inhibitory effect of osthole on alcohol-induced fatty liver in mice.

BACKGROUND: Alcohol is a major cause of fatty liver, the disease is a spectrum that is initiated with steatosis, and without therapy it is apt to develop inflammation, necrosis, fibrosis and finally cirrhosis. There are currently no ideal pharmacological reagents that can prevent or reverse this disease. Osthole is an active constituent isolated from the fruit of Cnidium monnieri (L.) Cusson, a Chinese herbal medicine, which has been used in clinics for many years. It has many functions such as anti-inflammation, anti-osteoporosis and anti-tumor and so on, but there is no report about treatment of alcoholic fatty liver in mice. AIM: To examine the inhibitory effect of osthole on alcohol-induced fatty liver in mice and to investigate the potential mechanisms. METHODS: A mouse model with alcoholic fatty liver was induced by orally feeding 52% erguotou wine by gavage when they were simultaneously treated with osthole 10, 20, 40 mg/kg for 4 weeks. Whereafter, the lipids in serum and hepatic tissue, the levels of malondialdehyde (MDA), superoxide dismutase (SOD), reduced glutathione hormone (GSH), tumor necrosis factor-alpha (TNF-alpha) in hepatic tissue, hepatic weight coefficient and its histological evaluation were measured. RESULTS: After treatment with osthole, the levels of serum total cholesterol (TC), triglyceride (TG), coefficient of hepatic weight, and the hepatic tissue contents of TC and TG were significantly decreased, the levels of MDA and TNF-alpha in liver were also decreased, while the GSH in liver was increased. Importantly, the histological evaluation of liver specimens demonstrated that osthole dramatically decreased lipid accumulation. CONCLUSION: Osthole could inhibit alcohol-induced fatty liver in mice, and the mechanism might be associated with its anti-oxidation and suppression of TNF-alpha production.

The tumour suppressor gene p53 modulates the severity of antigen-induced arthritis and the systemic immune response.

p53 is a transcription factor with a well-described role in the induction of apoptosis and cell cycle arrest as part of a protective response to a variety of stressful stimuli. Expansion of inflamed tissue in rheumatoid arthritis has been related to the loss of functioning p53, and the severity of collagen-induced arthritis is increased in p53-/- mice. Our objective was to assess the role of p53 in a model of adaptive immunity, antigen-induced arthritis (AIA). AIA was induced in p53-/- and wild-type mice by priming with methylated bovine serum albumin followed by intra-articular challenge. Severity of arthritis was assessed using a standardized scoring system and synovial apoptosis was detected by TdT-mediated biotin-dUTP nick-end labelling. Splenocyte proliferation was measured by [H(3)] incorporation and interferon (IFN)-gamma release. Splenocyte viability was assessed using Titreglow. Splenic T cell activation status was assessed by flow cytometry. Serum cytokines were measured using enzyme-linked immunosorbent assay. Increased severity of AIA in p53-/- mice was associated with decreased synovial apoptosis and with increased delayed-type hypersensitivity response, increased mitogen and antigen-induced splenocyte proliferation and increased IFN-gamma release in p53-/- mice compared with wild-type mice. Antigen-specific immunoglobulin responses were equivalent in both groups. Splenocyte viability was increased in p53-/- mice but T cell apoptosis was equivalent. T cell activation markers were increased in p53-/- mice compared with wild-type mice. Lipopolysaccharide-induced tumour necrosis factor release was increased in p53-/- mice with a trend to increased interleukin-6 in p53-/- mice compared with littermates. p53 is involved in the modulation of adaptive and innate immune responses relevant to arthritis models and is also involved in the modulation of severity of AIA by both cell-cycle dependent and cell-cycle-independent mechanisms.

Effect of placement of calcium sulphate when used for the repair of furcation perforations on the seal produced by a resin-based material.

AIM: To evaluate the sealing ability of calcium sulphate when used under composite resin for the repair of furcation perforations having different diameters. METHODOLOGY: Perforations of different diameter were created in the floors of pulp chambers in 60 extracted human molar teeth with either a number 3 (1 mm diameter) or 5 (1.5 mm diameter) round bur. The specimens of each group were divided into four sub-groups which were repaired with composite resin either alone or in combination with calcium sulphate that created an artificial floor (15 teeth group(-1)). Eight teeth without furcation perforations served as negative controls. In the leakage detection device, 1 mol L(-1) glucose solution was forced under a pressure of 1.5 KPa from the crown towards the pulp chamber floor. The concentration of leaked glucose was measured at 1, 2, 4, 7, 10, 15 and 20 days using a glucose oxidase method and the data evaluated using the rank sum test. RESULTS: The specimens with larger perforations repaired with composite resin alone had significantly more leakage (P < 0.05). Using calcium sulphate as an artificial floor significantly decreased leakage of smaller perforations (P < 0.05). In groups repaired with calcium sulphate under composite resin, leakage in smaller perforations was markedly lower than that in larger ones (P < 0.05). No significant difference was found between the specimens with 1 or 1.5 mm perforations repaired with resin alone (P > 0.05). CONCLUSIONS: Calcium sulphate significantly improved the sealing ability of 1 mm perforations repaired with composite resin but not for 1.5 mm perforations.