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1.  Visceral and somatic hypersensitivity, autonomic cardiovascular dysfunction and low-grade inflammation in a subset of irritable bowel syndrome patients*  
The pathophysiology of irritable bowel syndrome (IBS) is complex and not fully understood, so the aim of this study was to evaluate whether visceral and somatic hypersensitivity, autonomic cardiovascular dysfunction, and low-grade inflammation of the gut wall are associated with diarrhea-predominant IBS (D-IBS). Sixty-two patients with D-IBS and 20 control subjects participated in the study. Using the ascending method of limits (AML) protocol, we demonstrated that D-IBS patients had significantly lower sensory thresholds compared with healthy controls (P<0.001). Using diverse methods, especially the ischemic sensitivity test, for the first time in China, we confirmed that D-IBS patients have somatic hypersensitivity. They had a significantly higher systolic blood pressure and heart rate after a cold stimulus, indicative of autonomic cardiovascular dysfunction. Compared with the control group, D-IBS patients had a significantly higher level of calprotectin (P<0.001). We also found significant correlations between visceral and somatic hypersensitivity, visceral hypersensitivity and autonomic cardiovascular dysfunction, and somatic hypersensitivity and autonomic cardiovascular dysfunction. Our findings may provide valuable suggestions for the treatment of D-IBS.
PMCID: PMC4201319  PMID: 25294380
Irritable bowel syndrome (IBS); Visceral hypersensitivity; Somatic hypersensitivity; Autonomic cardiovascular dysfunction; Low-grade inflammation
2.  Spontaneous retroperitoneal hemorrhage after hemodialysis involving anticoagulant agents 
In this paper, we described the symptoms and treatment of a patient with diabetic nephropathy accompanied by spontaneous retroperitoneal hemorrhage after hemodialysis. An elderly female patient with diabetic nephropathy presented with severe pain, numbness, and an increasing swelling in the left hip and left thigh after six sessions of hemodialysis involving the use of an antiplatelet drug and an anticoagulant agent. Her hemoglobin decreased to 46 g/L. An abdominal ultrasound showed a hematoma in the left retroperitoneal space, and computed tomography (CT) findings revealed a 6 cm×8 cm×10 cm hematoma in the left psoas muscle. After aggressive supportive therapy [the administration of packed red blood cell transfusion, carbazochrome sodium sulfonate injection, and continuous venovenous hemofiltration (CVVH)], the patient’s vital signs stabilized and her hemoglobin increased to 86 g/L. Repeat CT showed that the hematoma had been partially absorbed after two weeks. Eventually, the patient was discharged with stable vital signs. Physicians should be aware of the possibility of spontaneous retroperitoneal hemorrhage, particularly in patients with diabetic nephropathy undergoing hemodialysis involving the use of anticoagulant agents.
PMCID: PMC3348233  PMID: 22556180
Spontaneous retroperitoneal hemorrhage; Diabetic nephropathy; Hemodialysis; Nadroparin calcium
3.  Production of conjugated linoleic acids by Lactobacillus plantarum strains isolated from naturally fermented Chinese pickles*  
Naturally fermented pickles harbour many lactic acid bacteria (LAB). Forty-three LAB strains with conjugated linoleic acid (CLA)-producing ability were isolated from three naturally fermented pickle brines. Of these isolates, lp15 identified as Lactobacillus plantarum by API 50 CHL system and full-length 16S rDNA sequence analysis exhibited the highest CLA-producing ability (26.1% conversion) at 48 h in de Man Rogosa Sharpe (MRS) broth in the presence of 100 µg/ml of linoleic acid (LA). Compared to other strains, L. plantarum strain lp15 showed the highest tolerance upon increased levels of LA in the medium, i.e., up to 600 µg/ml. This strain converted about 25% of LA into CLA isomers [predominantly cis-9, trans-11 CLA (9-CLA) and trans-10, cis-12 CLA (10-CLA)], of which 75% was 9-CLA. Interestingly, though the conversion rate of LA into CLA by lp15 remained stable between 100 to 600 µg/ml LA levels in the medium, it dropped sharply at 1000 µg/ml. Taken together, the lp15 strain displayed relatively high LA tolerance with higher conversion rate, which implies that this strain is a valuable candidate for enhancing the CLA content in food-sources like pickles.
PMCID: PMC3208172  PMID: 22042657
Conjugated linoleic acids; Lactobacillus plantarum; Lactic acid bacteria; Pickle; Gas chromatography
4.  Development and evaluation of immunoassay for zeranol in bovine urine*  
A high affinity polyclonal antibody-based enzyme linked immunosorbent assay (ELISA) was developed for the quantification of zeranol in bovine urine. On the basis of urine matrix studies, the optimized dilution factors producing insignificant matrix interference were selected as 1:5 in pretreatment. In the improved ELISA, the linear response range was between 0.02 and 1 μg/ml , and the detection limit was 0.02 μg/ml for the assay. The overall recoveries and the coefficients of variation (CVs) were in the range of 82%~127% and 3.5%~8.8%, respectively. Thirty-six bovine urine samples spiked with zeranol (ranging from 0.2 to 10 μg/ml) were detected by the ELISA and liquid chromatography (LC) method, and good correlations were obtained between the two methods (R 2=0.9643). We conclude that this improved ELISA is suitable tool for a mass zeranol screening and can be an alternative for the conventional LC method for zeranol in bovine urine.
PMCID: PMC2100163  PMID: 18257125
Zeranol; Enzyme linked immunosorbent assay (ELISA); Bovine urine
5.  Zirconium oxide ceramic foam: a promising supporting biomaterial for massive production of glial cell line-derived neurotrophic factor*  
This study investigated the potential application of a zirconium oxide (ZrO2) ceramic foam culturing system to the production of glial cell line-derived neurotrophic factor (GDNF). Three sets of ZrO2 ceramic foams with different pore densities of 10, 20, and 30 pores per linear inch (PPI) were prepared to support a 3D culturing system. After primary astrocytes were cultured in these systems, production yields of GDNF were evaluated. The biomaterial biocompatibility, cell proliferation and activation of cellular signaling pathways in GDNF synthesis and secretion in the culturing systems were also assessed and compared with a conventional culturing system. In this study, we found that the ZrO2 ceramic foam culturing system was biocompatible, using which the GDNF yields were elevated and sustained by stimulated cell proliferation and activation of signaling pathways in astrocytes cultured in the system. In conclusion, the ZrO2 ceramic foam is promising for the development of a GDNF mass production device for Parkinson’s disease treatment.
PMCID: PMC4265555  PMID: 25471830
Zirconium oxide; Ceramic foam; Glial cell line-derived neurotrophic factor (GDNF); Parkinson’s disease
6.  Influence of heat stress on leaf ultrastructure, photosynthetic performance, and ascorbate peroxidase gene expression of two pear cultivars (Pyrus pyrifolia)*  
Plants encounter a variety of stresses in natural environments. One-year-old pot-grown trees of pear (Pyrus pyrifolia Nakai cv. Cuiguan and Wonhwang) were exposed to two heat stress regimes. Under constant short-term heat stress, chloroplasts and mitochondria were visibly damaged. Relative chlorophyll content and maximum photochemical efficiency of photosystem II were significantly decreased, which indicated that the leaf photosynthetic capability declined. Under chronic heat stress, mesophyll cell ultrastructure was not obviously damaged, but leaf photosynthetic capability was still restrained. As chronic heat stress was a simulation of the natural environment in summer, further study of the responses under this stress regime was undertaken. Ascorbate peroxidase (APX) activity was increased in ‘Cuiguan’, but not in ‘Wonhwang’. Inducible expression of PpAPX genes in the cytoplasm, chloroplasts and peroxisomes was consistent with increased APX activity in ‘Cuiguan’, whereas only weak induction of PpAPX genes was observed in ‘Wonhwang’. The isoenzymes cytosolic APX1 (cAPX1) and stromal APX (sAPX) were confirmed to be localized in the cytoplasm and chloroplasts, respectively.
PMCID: PMC3863366  PMID: 24302708
Pear; Ultrastructure; Ascorbate peroxidase; Subcellular localization; Synergistic effect
7.  Cloning and functional analysis of a novel ascorbate peroxidase (APX) gene from Anthurium andraeanum *  
An 888-bp full-length ascorbate peroxidase (APX) complementary DNA (cDNA) gene was cloned from Anthurium andraeanum, and designated as AnAPX. It contains a 110-bp 5′-noncoding region, a 28-bp 3′-noncoding region, and a 750-bp open reading frame (ORF). This protein is hydrophilic with an aliphatic index of 81.64 and its structure consisting of α-helixes, β-turns, and random coils. The AnAPX protein showed 93%, 87%, 87%, 87%, and 86% similarities to the APX homologs from Zantedeschia aethiopica, Vitis pseudoreticulata, Gossypium hirsutum, Elaeis guineensis, and Zea mays, respectively. AnAPX gene transcript was measured non-significantly in roots, stems, leaves, spathes, and spadices by real-time polymerase chain reaction (RT-PCR) analysis. Interestingly, this gene expression was remarkably up-regulated in response to a cold stress under 6 °C, implying that AnAPX might play an important role in A. andraeanum tolerance to cold stress. To confirm this function we overexpressed AnAPX in tobacco plants by transformation with an AnAPX expression construct driven by CaMV 35S promoter. The transformed tobacco seedlings under 4 °C showed less electrolyte leakage (EL) and malondialdehyde (MDA) content than the control. The content of MDA was correlated with chilling tolerance in these transgenic plants. These results show that AnAPX can prevent the chilling challenged plant from cell membrane damage and ultimately enhance the plant cold tolerance.
PMCID: PMC3863369  PMID: 24302711
AnAPX; Gene expression; Cold stress; Anthurium andraeanum
8.  Correlation between virus persistent infection and cardic function in patients with dilated cardiomyopathy*  
In our study, 50 patients with dilated cardiomyopathy (DCM) were selected to investigate the correlation between virus persistent infection and cardic function. We found that 44% of patients with DCM were coxsackie virus B-RNA (CVB-RNA) positive, significantly different from that (20%) of the normal control group (P<0.05). The expression levels of coxsackie adenovirus receptor (CAR) in patients with DCM were significantly higher than those in the normal control group (P<0.01). In CVB-RNA-positive patients, expression levels of CAR were significantly higher than those in CVB-RNA-negative patients (P<0.01). There was a positive correlation between CAR expression and brain natriuretic peptide (BNP) level in patients with DCM, but no significant correlations between the CAR expression level and left ventricular ejection fraction (LVEF) or left ventricular end diastolic diameter (LVEDd). These results showed that expression levels of CAR on the surface of white cells can be used as an indicator for detecting persistent virus infection. We found that expression levels of CAR and heart function in patients with DCM were highly correlated.
PMCID: PMC3735976  PMID: 23897795
9.  A survey of five first-level hospital ethics committees in Urumqi, China*  
This paper presents the results from a survey of first-level hospitals in Urumqi, China. The survey had two parts: the first part was aimed at understanding the operation of the ethics committees of first-level hospitals, including the process for electing members and the variety of members’ backgrounds. Information was also gathered about the establishment of criteria, operational rules and regulations, and standard operational procedures. The aim of the second part was to investigate the level of understanding among technicians and doctors about the function of the ethics committees. This paper identifies and analyzes some deficiencies found in the operation of hospital ethics committees, offers some constructive suggestions for improvement, and promotes the role of the Xinjiang Uygur autonomous region hospital ethics committees.
PMCID: PMC3682171  PMID: 23733432
First-level hospital; Ethics committee; Xinjiang Urumqi
10.  Association between mandibular posterior alveolar morphology and growth pattern in a Chinese population with normal occlusion*  
Objective: To investigate the relationship between growth patterns and mandibular posterior tooth-alveolar bone complex morphology in a Chinese population with normal occlusion. Methods: Forty-five patients with normal occlusion (23 males, 22 females) were included in this study. Among these patients, 20 displayed the vertical growth pattern, and 20 had the horizontal growth pattern, while the remaining patients displayed the average growth pattern. All of the patients underwent dental cone beam computed tomography (CBCT), which included the region of the mandibular posterior teeth and the alveolar. A linear regression analysis and a correlation analysis between the facial height index (FHI) and the alveolar bone morphology were performed. Results: The inclination of the molars, the thickness of the cortical bone, and the height of the mandibular bone differed significantly between patients with the horizontal growth pattern and those with the vertical growth pattern (P<0.05). Significant positive correlations were found between: the FHI and the inclination of the molars; the FHI and the thickness of the cortical bone; and the FHI and the height of the mandibular bone. Conclusions: The mandibular posterior tooth-alveolar bone complex morphology may be affected by growth patterns.
PMCID: PMC3542955  PMID: 23303628
Cone beam computed tomography (CBCT); Growth pattern; Alveolar morphology; Normal occlusion
11.  Establishment of reference mandibular plane for anterior alveolar morphology evaluation using cone beam computed tomography*  
To propose a method of establishing the reference mandibular plane (MP), which could be reestablished according to the coordinates of the reference points, and then facilitate the assessment of anterior alveolar morphology using cone beam computed tomography (CBCT), sixty patients with bimaxillary protrusion were randomly selected and CBCT scans were taken. The CBCT scans were transferred to Materialism’s interactive medical image control system 10.01 (MIMICS 10.01), and three dimensional models of the entire jaws were constructed. Reference points determining the reference MP were positioned in the coronal, axial, sagittal windows, and the points were exactly located by recording their coordinates in the interfaces of software. The reference MP provided high intra-observer reliability (Pearson’s r 0.992 to 0.999), and inter-observer reliability (intra-class correlation coefficients (ICCs) 0.996 to 0.999).
PMCID: PMC3494033  PMID: 23125087
Reference Plane; Cone beam computed tomography; Alveolar morphology
12.  Recent developments in the detection of melamine*  
In recent years, there were two reported outbreaks of food borne illness associated with melamine. The presence of melamine and its related compounds in milk, feed, and other foods has resulted in the need for reliable methods for the detection and accurate quantification of this class of contaminants. The sample pretreatment for melamine in a complex matrix usually involves a liquid extraction by a polar solvent, followed by a further clean-up with solid phase extraction. Analyses of melamine and related compounds are commonly carried out by liquid or gas chromatographic methods conjugated with mass spectrometry. Other innovative screening methods, which use antibodies, molecularly imprinted polymers, capillary electrophoresis, and gold nanoparticles, are also used to develop assays and biosensors to melamine. However, many of these methods have been hindered by matrix effects, the solubility of melamine-cyanuric acid complex, and background contamination. This article reviews recent developments for detecting melamine and discusses future directions.
PMCID: PMC3390710  PMID: 22761244
Melamine; Detection; Confirmation methods; Screening methods; Sample pretreatment
13.  Development of an electrochemical immunoassay for detection of gatifloxacin in swine urine*  
To detect gatifloxacin (GAT) residue in swine urine, an electrochemical immunoassay was established. An indirect competitive immunoassay was developed, in which the coating antigen is immobilized in an enzyme-linked immunosorbent assay (ELISA) plate and GAT residue from the sample competes with the limited binding sites in added anti-GAT antibody. Horseradish peroxidase (HRP) conjugated to goat anti-rabbit IgG was used as the enzymatic label. A carbon fiber working electrode was constructed and current signals were detected by using hydrogen peroxide as a substrate and hydroquinone as an electrochemical mediator. The electrochemical immunoassay was evaluated by analysis of GAT in buffer or swine urine and an average value of half inhibition concentration (IC50) of 8.9 ng/ml was obtained. Excellent specificity of the antibody was achieved with little cross-reaction with lomefloxacin (3.0%), ciprofloxacin (3.0%), and ofloxacin (1.9%) among commonly used (fluoro)quinolones. In conclusion, the immunoassay system developed in this research can be used as a rapid, powerful and on-site analytical tool to detect GAT residue in foods and food products.
PMCID: PMC3274739  PMID: 22302425
Gatifloxacin; Drug residue; Antibody; Carbon fiber electrode; Electrochemical immunoassay
14.  Three-dimensional evaluation of upper anterior alveolar bone dehiscence after incisor retraction and intrusion in adult patients with bimaxillary protrusion malocclusion*  
Objective: The purpose of this study was to evaluate three-dimensional (3D) dehiscence of upper anterior alveolar bone during incisor retraction and intrusion in adult patients with maximum anchorage. Methods: Twenty adult patients with bimaxillary dentoalveolar protrusion had the four first premolars extracted. Miniscrews were placed to provide maximum anchorage for upper incisor retraction and intrusion. A computed tomography (CT) scan was performed after placement of the miniscrews and treatment. The 3D reconstructions of pre- and post-CT data were used to assess the dehiscence of upper anterior alveolar bone. Results: The amounts of upper incisor retraction at the edge and apex were (7.64±1.68) and (3.91±2.10) mm, respectively, and (1.34±0.74) mm of upper central incisor intrusion. Upper alveolar bone height losses at labial alveolar ridge crest (LAC) and palatal alveolar ridge crest (PAC) were 0.543 and 2.612 mm, respectively, and the percentages were (6.49±3.54)% and (27.42±9.77)%, respectively. The shape deformations of LAC-labial cortex bending point (LBP) and PAC-palatal cortex bending point (PBP) were (15.37±5.20)° and (6.43±3.27)°, respectively. Conclusions: Thus, for adult patients with bimaxillary protrusion, mechanobiological response of anterior alveolus should be taken into account during incisor retraction and intrusion. Pursuit of maximum anchorage might lead to upper anterior alveolar bone loss.
PMCID: PMC3232432  PMID: 22135148
Alveolar bone loss; Adult patients; Computed tomography; Three-dimensional registration
15.  Effect of dietary fatty acids on serum parameters, fatty acid compositions, and liver histology in Shaoxing laying ducks*  
The effects of different fatty acid (FA) contents in diet on serum parameters, FA compositions of eggs and meat, and liver morphological changes were studied in Shaoxing laying ducks. A total of 264 ducks at 17 weeks were fed a control diet or a diet containing 30 g/kg fish oil (FO), 25 g/kg sunflower oil (SO), or 30 g/kg palm oil with 20 g/kg beef tallow (PBO). Malondialdehyde (MDA) content in the liver and the serum of ducks fed the PBO diet was significantly (P<0.05) higher than that of ducks fed the other diets. Triglyceride (TG) and total cholesterol (TC) levels were significantly lower (P<0.05) in ducks fed the FO diet. Serum TC also was lower in ducks fed the SO diet. Superoxide dismutase (SOD) activity was also affected by diets. The contents of polyunsaturated FAs (PUFAs) in eggs and meat were significantly higher (P<0.001) in ducks fed the FO and SO diets than in ducks fed the control diet. The level of C22:6 (n-3) FA in ducks fed the FO diet was significantly higher than that in ducks fed the other diets. However, the conversion efficiency of the longer-chain C20:5 (n-3) FA was higher than that of C22:6 (n-3). Ducks fed the PBO diet exhibited lipid droplet accumulation in the liver. These results demonstrate that a diet enriched with different FAs has strong effects on serum lipid levels and the deposition of PUFAs into tissue lipids.
PMCID: PMC3167907  PMID: 21887849
Duck; Liver; Egg; Meat; Fatty acid; Lipid oxidation
16.  Kinetic analysis of γ-glutamyltransferase reaction process for measuring activity via an integration strategy at low concentrations of γ-glutamyl p-nitroaniline*  
At 0.12 mmol/L γ-glutamyl p-nitroaniline (GGPNA), an improved integrated method was developed for kinetic analysis of γ-glutamyltransferase (GGT) reaction process and the integration with the classical initial rate method to measure serum GGT. For the improved integrated method, an integrated rate equation, which used the predictor variable of reaction time and considered inhibitions by both GGPNA and products, was nonlinearly fit to GGT reaction processes. For the integration strategy, classical initial rates were estimated when GGPNA consumption percentages were below 50%; otherwise, maximal reaction rates of GGT were estimated by the improved integrated method and converted into initial rates according to the differential rate equation at 0.11 mmol/L GGPNA. The integration strategy was validated using optimized GGT kinetic parameters and 10-s intervals to record reaction curves within 8.0 min. By the integration strategy, there was a linear response from 0.9 to 32.0 U/L GGT, coefficients of variation were below 3.5% for GGT from 8.0 to 32.0 U/L (n=5), and GGT activities in clinical sera responded linearly to their classical initial rates at 2.00 mmol/L GGPNA with an expected slope. Therefore, the integration strategy was successful in measuring GGT at 0.12 mmol/L GGPNA.
PMCID: PMC3048932  PMID: 21370502
Integration strategy; Chromogenic substrate; Data processing; γ-Glutamyltransferase; Kinetic analysis; Serum enzyme assay
17.  A food-grade industrial arming yeast expressing β-1,3-1,4-glucanase with enhanced thermal stability*  
The aim of this work was to construct a novel food-grade industrial arming yeast displaying β-1,3-1,4-glucanase and to evaluate the thermal stability of the glucanase for practical application. For this purpose, a bi-directional vector containing galactokinase (GAL1) and phosphoglycerate kinase 1 (PGK1) promoters in different orientations was constructed. The β-1,3-1,4-glucanase gene from Bacillus subtilis was fused to α-agglutinin and expressed under the control of the GAL1 promoter. α-galactosidase induced by the constitutive PGK1 promoter was used as a food-grade selection marker. The feasibility of the α-galactosidase marker was confirmed by the growth of transformants harboring the constructed vector on a medium containing melibiose as a sole carbon source, and by the clear halo around the transformants in Congo-red plates owing to the expression of β-1,3-1,4-glucanase. The analysis of β-1,3-1,4-glucanase activity in cell pellets and in the supernatant of the recombinant yeast strain revealed that β-1,3-1,4-glucanase was successfully displayed on the cell surface of the yeast. The displayed β-1,3-1,4-glucanase activity in the recombinant yeast cells increased immediately after the addition of galactose and reached 45.1 U/ml after 32-h induction. The thermal stability of β-1,3-1,4-glucanase displayed in the recombinant yeast cells was enhanced compared with the free enzyme. These results suggest that the constructed food-grade yeast has the potential to improve the brewing properties of beer.
PMCID: PMC2801089  PMID: 20043351
α-agglutinin; Food-grade selection marker; β-1,3-1,4-glucanase; α-galactosidase; Thermostability
18.  Viral infection of tobacco plants improves performance of Bemisia tabaci but more so for an invasive than for an indigenous biotype of the whitefly*  
The ecological effects of plant-virus-vector interactions on invasion of alien plant viral vectors have been rarely investigated. We examined the transmission of Tomato yellow leaf curl China virus (TYLCCNV) by the invasive Q biotype and the indigenous ZHJ2 biotype of the whitefly Bemisia tabaci, a plant viral vector, as well as the influence of TYLCCNV-infection of plants on the performance of the two whitefly biotypes. Both whitefly biotypes were able to acquire viruses from infected plants and retained them in their bodies, but were unable to transmit them to either tobacco or tomato plants. However, when the Q biotype fed on tobacco plants infected with TYLCCNV, its fecundity and longevity were increased by 7- and 1-fold, respectively, compared to those of the Q biotype fed on uninfected tobacco plants. When the ZHJ2 biotype fed on virus-infected plants, its fecundity and longevity were increased by only 2- and 0.5-fold, respectively. These data show that the Q biotype acquired higher beneficial effects from TYLCCNV-infection of tobacco plants than the ZHJ2 biotype. Thus, the Q biotype whitefly may have advantages in its invasion and displacement of the indigenous ZHJ2 biotype.
PMCID: PMC2801088  PMID: 20043350
Bemisia tabaci; Biotypes; Begomovirus; Vector-virus interaction; Biological invasion
19.  Study on swelling model and thermodynamic structure of native konjac glucomannan 
We investigated the higher structure of konjac glucomannan (KGM) in the amorphous state and solution using a laser particle size analyzer and a water activity meter. The results show that the thermodynamic structures of native KGM were primarily composed of the lamella structure units, which involve both granular crystalline and amorphous regions, and that the connection zones of such units contained both loose and tight aggregation regions. The value of surface tension (σ) of native KGM, resting with the density of its hydroxyl groups’ self-association, was an important parameter to analyze the higher structures of native KGM in the thermodynamic swelling model of native KGM.
PMCID: PMC2666203  PMID: 19353745
Swelling model; Thermodynamic structure; Konjac glucomannan (KGM); Higher structure
20.  L-Proline as an efficient and reusable promoter for the synthesis of coumarins in ionic liquid 
The effect of L-proline as a promoter on the condensation reaction of salicylaldehyde or its derivatives with ethyl acetoacetate in neutral ionic liquid [emim]BF4 was studied. All reactions were carried out under mild reaction conditions and achieved high yields. Moreover, the ionic liquid containing L-proline could be recycled and reused for several times without noticeably decreasing in productivity. The results show that the L-proline-[emim]BF4 system has a potential in contribution to the development of environmentally friendly and inexpensive processes in organic syntheses.
PMCID: PMC2596292  PMID: 19067468
L-Proline; Ionic liquid; Coumarins; Knoevenagel condensation
21.  A simple and effective method for total RNA isolation of appressoria in Magnaporthe oryzae * §  
Appressorium formation is an important event in establishing a successful interaction between the rice blast fungus, Magnaporthe oryzae, and its host plant, rice. An understanding of molecular events occurring in appressorium differentiation will give new strategies to control rice blast. A quick and reliable method to extract total RNA from appressorium is essential for studying gene expression during appressorium formation and its mechanism. We found that duplicate film is an efficient substratum for appressorium formation, even when inoculated with high density conidia. When inoculated with conidia at 1×106 ml−1, the percentages of conidium germination and appressorium formation were (97.98±0.67)% and (97.88±0.45)%, respectively. We applied Trizol before appressorium collection for total RNA isolation, and as much as 113.6 μg total RNA was isolated from the mature appressoria at 24 h after inoculation. Functional analysis of two genes, MNH6 and MgATG1, isolated from the cDNA subtractive library, revealed that the quantity of RNA was good enough to construct a cDNA (complementary DNA) library or a cDNA subtractive library. This method may be also applicable for the appressorium RNA isolation of other pathogenic fungi in which conidia differentiate into appressoria in the early stages of host infection.
PMCID: PMC2565745  PMID: 18837109
Appressorium; Magnaporthe oryzae; RNA isolation
22.  Effects of over-expressing resistin on glucose and lipid metabolism in mice*  
Resistin, a newly discovered peptide hormone mainly secreted by adipose tissues, is present at high levels in serum of obese mice and may be a potential link between obesity and insulin resistance in rodents. However, some studies of rat and mouse models have associated insulin resistance and obesity with decreased resistin expression. In humans, no relationship between resistin level and insulin resistance or adiposity was observed. This suggests that additional studies are necessary to determine the specific role of resistin in the regulation of energy metabolism and adipogenesis. In the present study, we investigated the effect of resistin in vivo on glucose and lipid metabolism by over-expressing resistin in mice by intramuscular injection of a recombinant eukaryotic expression vector pcDNA3.1-Retn encoding porcine resistin gene. After injection, serum resistin and serum glucose (GLU) levels were significantly increased in the pcDNA3.1-Retn-treated mice; there was an obvious difference in total cholesterol (TC) level between the experiment and the control groups on Day 30. In pcDNA3.1-Retn-treated mice, both free fatty acid (FFA) and high density lipoprotein (HDL) cholesterol levels were markedly lower than those of control, whereas HDL cholesterol and triglyceride (TG) levels did not differ between the two groups. Furthermore, lipase activity was expressly lower on Day 20. Our data suggest that resistin over-expressed in mice might be responsible for insulin resistance and parameters related to glucose and lipid metabolism were changed accordingly.
PMCID: PMC2170468  PMID: 18196612
Resistin; Glucose; Lipid; Metabolism
23.  Lance-Adams syndrome: a report of two cases*  
Chronic post-hypoxic myoclonus, also known as Lance-Adams syndrome (LAS), is a rare complication of successful cardiopulmanry resuscitation often accompanied by action myoclonus and cerebellar ataxia. It is seen in patients who have undergone a cardiorespiratory arrest, regained consciousness afterwards, and then developed myoclonus days or weeks after the event. Worldwide, 122 cases have been reported in the literature so far, including 1 case of Chinese. Here we report 2 Chinese LAS patients with detailed neuroimagings. Cranial single photon emission computed tomography (SPECT) of patient 1, a 52-year-old woman, showed a mild hypoperfusion in her left temporal lobe, whereas patient 2, a 54-year-old woman, manifested a mild bilateral decrease of glucose metabolism in the frontal lobes and a mild to moderate decrease of the N-acetyl aspartate (NAA) peak in the bilateral hippocampi by cranial [18F]-fluorodeoxyglucose positron emission tomographic (PET) scan and cranial magnetic resonance spectroscopy (MRS), respectively. We also review the literature on the neuroimaging, pathogenesis, and treatment of LAS.
PMCID: PMC1997224  PMID: 17910113
Lance-Adams syndrome; Chronic post-hypoxic myoclonus; Action myoclonus; Cerebellar ataxia; Single photon emission computed tomography; Positron emission tomography; Magnetic resonance spectroscopy
24.  Lead accumulation and tolerance of Moso bamboo (Phyllostachys pubescens) seedlings: applications of phytoremediation*  
A hydroponics experiment was aimed at identifying the lead (Pb) tolerance and phytoremediation potential of Moso bamboo (Phyllostachys pubescens) seedlings grown under different Pb treatments. Experimental results indicated that at the highest Pb concentration (400 μmol/L), the growth of bamboo seedlings was inhibited and Pb concentrations in leaves, stems, and roots reached the maximum of 148.8, 482.2, and 4282.8 mg/kg, respectively. Scanning electron microscopy revealed that the excessive Pb caused decreased stomatal opening, formation of abundant inclusions in roots, and just a few inclusions in stems. The ultrastructural analysis using transmission electron microscopy revealed that the addition of excessive Pb caused abnormally shaped chloroplasts, disappearance of endoplasmic reticulum, shrinkage of nucleus and nucleolus, and loss of thylakoid membranes. Although ultrastructural analysis revealed some internal damage, even the plants exposed to 400 μmol/L Pb survived and no visual Pb toxicity symptoms such as necrosis and chlorosis were observed in these plants. Even at the highest Pb treatment, no significant difference was observed for the dry weight of stem compared with controls. It is suggested that use of Moso bamboo as an experimental material provides a new perspective for remediation of heavy metal contaminated soil owing to its high metal tolerance and greater biomass.
PMCID: PMC4322423  PMID: 25644467
Moso bamboo; Pb; Phytoremediation; Scanning electron microscopy; Transmission electron microscopy
25.  Determination of royal jelly freshness by ELISA with a highly specific anti-apalbumin 1, major royal jelly protein 1 antibody*  
Major royal jelly protein 1 (MRJP1), designated apalbumin 1, has been regarded as a freshness marker of royal jelly (RJ). A MRJP1-specific peptide (IKEALPHVPIFD) identified by bioinformatics analysis of homologous members of the major royal protein family was synthesized and used to raise polyclonal anti-MRJP1 antibody (anti-SP-MRJP1 antibody). Western blot analysis showed that anti-SP-MRJP1 antibody only reacted with MRJP1 in RJ. In contrast, the previously reported antibody against recombinant MRJP1 (anti-R-MRJP1 antibody) reacted with other members of MRJP family in RJ. Enzyme-linked immunosorbent assay (ELISA) using anti-SP-MRJP1 antibody demonstrated that MRJP1 content in RJ stored at 40 °C significantly degraded by 37.3%, 55.9%, 58.0%, 60.6%, 65.7%, 72.7%, and 73.1% at 7, 14, 21, 28, 35, 42, and 49 d, respectively, when compared with MRJP1 content in fresh RJ (0 d). Optical density analysis of MRJP bands from sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) profiles demonstrated that the degradation of MRJP1, MRJP2, MRJP3, and MRJP5 in RJ was strongly and positively correlated with the period of storage (P<0.0001). Our results indicated anti-SP-MRJP1 antibody was highly specific for MRJP1, and ELISA using the antibody is a sensitive and easy-to-use method to determine the freshness and authenticity of RJ.
PMCID: PMC4322426  PMID: 25644470
Freshness; Royal jelly; Major royal jelly protein 1 (MRJP1); Enzyme-linked immunosorbent assay (ELISA); High specific antibody

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