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Core-shell structural adenosine-imprinted microspheres were prepared via a two-step procedure. Polystyrene core particles (CP) were firstly prepared via a reversible addition-fragmentation chain transfer (RAFT) polymerization leaving the iniferter on the surface of the cores, then a molecularly imprinted polymer (MIP) shell was synthesized on the surface of the cores by using acrylamide (AAm) as the functional monomer and ethylene glycol dimethacrylate (EGDMA) as the cross-linker. The formation and growth of the MIP layer were seen dependent on the initiator (AIBN), AAm and the polymerization time used within the polymerization. SEM/TEM images showed that the dimensions of the cores and shells were 2μM and 44nm, respectively. The MIP microspheres exhibited a fast rebinding rate within 2h and a maximum adsorption capacity of 177μg per gram for adenosine. The adsorption fitted a Langmuir-Freundlich (LF) isotherm model with a K LF value of 41mL/μg and a q m value of 177μg/g for the MIP microspheres. The values were larger than those for a non-molecularly imprinted polymer (NIP) particles (5mL/μg and 88μg/g) indicating a better adsorption ability towards adenosine. The MIP microspheres showed a good selectivity for adenosine with a higher adsorption (683nmol/g) for adenosine than that (91nmol/g, 24nmol/g and 54nmol/g) for guanosine, cytidine and uridine respectively. Further experiment proved that the adenosine-imprinted polymer microspheres also had a good selectivity for ADP-ribosylated proteins that the MIP could extract the ADP-ribosylated proteins from the cell extract samples., (Copyright © 2016 Elsevier B.V. All rights reserved.)

To investigate the antitumor effect of anthocyanins extracted from Chinese bayberry fruit ( Myrica rubra Sieb. et Zucc.), a nude mouse tumor xenograft model was established. Treatments with C3G (cyanidin-3-glucoside, an anthocyanin) significantly suppressed the growth of SGC-7901 tumor xenografts in a dose-dependent manner. Immunohistochemical staining showed a significant increase in p21 expression, indicating that the cell cycle of tumor xenografts was inhibited. qPCR screening showed that C3G treatment up-regulated the expression of the KLF6 gene, which is an important tumor suppressor gene inactivated in many human cancers. Western blot showed that C3G treatments markedly increased KLF6 and p21 protein levels, inhibited CDK4 and Cyclin D1 expression, but did not notably change the expression of p53. These results indicated that KLF6 up-regulates p21 in a p53-independent manner and significantly reduces tumor proliferation. This study provides important information for the possible mechanism of C3G-induced antitumor activity against gastric adenocarcinoma in vivo., Competing Interests: The authors declare no conflict of interest.

This retrospective study used a population-based national registry to determine the impact of local treatment modalities on survival in patients with metastatic esophageal cancer (EC). The Surveillance Epidemiology and End Results (SEER) database was used to identify patients with metastatic EC from 1988 to 2012. A total of 9,125 patients were identified. There were 426 patients underwent primary surgery, 4,786 patients were administered radiotherapy (RT) alone, 847 patients underwent surgery plus RT, and 3,066 patients without any local treatment. Multivariate analysis results indicated that year of diagnosis, age, race, histologic subtype, grade, and local treatment modalities were independent prognostic factors for overall survival (OS). The 5-year OS were 8.4%, 4.5%, 17.5%, and 3.4% in primary surgery, RT only, surgery plus RT, and no local treatment, respectively (P < 0.001). Subgroup analyses showed that the impact of RT was mainly reflected by preoperative radiotherapy, as patients received preoperative radiotherapy had significantly better OS than patients who underwent primary surgery alone and postoperative RT, the 5-year OS rates were 24.7%, 6.5%, and 7.8%, respectively, respectively (P < 0.001). Surgery plus RT, especially preoperative RT, may improve long-term survival of patients with metastatic EC.

Objective: To investigate the biodistribution and single-photon emission computed tomography (SPECT) imaging of (99m)Tc-labeled arginine-glutamic acid-threonine (RET) and arginine-glutamic acid-glycine (REG) in nude mice bearing human lung cancer xenografts., Materials and Methods: RET and REG were labeled directly with (99m)Tc and their binding efficiency to tumor cells was measured in human nonsmall cell lung cancer H1299 cells. After intravenously injecting (99m)Tc-RET and (99m)Tc-REG into normal mice and nude mice bearing human lung cancer xenografts, their biodistribution was measured at different postinjection times, and percentages of injected dose per gram tissue (% ID/g) of organs of interest were calculated. The mice bearing H1299 lung cancer xenografts were scanned by SPECT at different times following the (99m)Tc-RET or (99m)Tc-REG injection., Results: The radiochemical purity of (99m)Tc-RET and (99m)Tc-REG was 93.15%±2.02% and 92.90%±2.86%, respectively. The binding rate of (99m)Tc-RET and (99m)Tc-REG to H1299 cells was 3.56%±0.37% and 2.32%±0.31%, respectively. The uptake of (99m)Tc-RET and (99m)Tc-REG in tumor was 4.96±1.05% ID/g at 4 hours postinjection and 1.95±0.73% ID/g at 2 hours postinjection, respectively. Tumors in nude mice could be best imaged at 4.5-6 hours postinjection of (99m)Tc-RET., Conclusion: (99m)Tc-RET has a higher binding rate to H1299 cells than (99m)Tc-REG and might be used as a potential lung cancer imaging agent.

HNO has recently been found to possess distinct biological functions from NO. Studying the biological functions of HNO calls for the development of sensitive and selective fluorescent probes. Herein, we designed and synthesized a FRET-based ratiometric probe to detect HNO in living cells. Our studies revealed that the probe is capable of detecting HNO in a rapid and ratiometric manner under physiological conditions. In bioimaging studies, the probe displayed a clear color change from blue to green when treated with HNO.

Three-dimensional surface wrinkling of soft cylindrical tissues induced by differential growth is explored. Differential volumetric growth can cause their morphological stability, leading to the formation of hexagonal and labyrinth wrinkles. During postbuckling, multiple bifurcations and morphological transitions may occur as a consequence of continuous growth in the surface layer. The physical mechanisms underpinning the morphological evolution are examined from the viewpoint of energy. Surface curvature is found to play a regulatory role in the pattern evolution. This study may not only help understand the morphogenesis of soft biological tissues, but also inspire novel routes for creating desired surface patterns of soft materials.

Plastids are ideal subcellular hosts for the expression of transgenes and have been successfully used for the production of different biopolymers, therapeutic proteins and industrial enzymes. Phaeodactylum tricornutum is a widely used aquatic feed species. In this study, we focused on developing a high-efficiency plastid expression system for P. tricornutum. In the plastid transformation vector, the site selected for integration was the transcriptionally active intergenic region present between the trnI and trnA genes, located in the IR (inverted repeat) regions of the plastid genome. Initially, a CAT reporter gene (encoding chloramphenicol acetyltransferase) was integrated at this site in the plastid genome. The expression of CAT in the transformed microalgae conferred resistance to the antibiotic chloramphenicol, which enabled growth in the selection media. Overall, the plastid transformation efficiency was found to be approximately one transplastomic colony per 1,000 microalgae cells. Subsequently, a heterologous gene expression cassette for high-level expression of the target gene was created and cloned between the homologous recombination elements. A TA cloning strategy based on the designed XcmI-XcmI sites could conveniently clone the heterologous gene. An eGFP (green fluorescent protein) reporter gene was used to test the expression level in the plastid system. The relatively high-level expression of eGFP without codon optimisation in stably transformed microalgae was determined to account for 0.12 % of the total soluble protein. Thus, this study presents the first and convenient plastid gene expression system for diatoms and represents an interesting tool to study diatom plastids.

Methyl parathion hydrolase (MPH), an enzyme that catalyses the turnover of methyl parathion (MP) to p-nitrophenol (pNP), can be utilized as an enzyme label. In this paper, a unique fluorescence response of 1,1-bis[4-(diethylaminomethyl)phenyl]-2,3,4,5-tetraphenylsilole (A 2 HPS) to MPH whose gene was obtained from Pseudomonas sp. strain WBC-3 is described. In the absence of MP, A 2 HPS could only give a small fluorescence response to the enzyme (I/I 0 = 1.1). The detection must be performed under low pH conditions, and the influence of BSA and hemoglobin (Hb) was high; upon addition of the enzyme's substrate, 1 × 10 -5 μg mL -1 or 2.85 × 10 -13 M of the MPH could be reported by A 2 HPS with a higher I/I 0 of 1.7. The detection limit was 10 5 times more sensitive than that given by a spectrophotometric method. In addition, the assay could be performed at a near neutral pH which was more biocompatible, and little influence was observed from BSA and Hb. The light-on response to the MPH was due to the different quenching effect of the MP and pNP on A 2 HPS and the improvement in the detection selectivity was due to combining the enzyme reaction with the detection. The findings of this work suggested that A 2 HPS and MP could form a new reporter system for the MPH enzyme label.

Neurological degeneration can occur after compression of the spinal cord. It is widely accepted that spinal cord compression leads to ischemic lesions and ultimately neurological dysfunction due to a narrowed spinal canal. Therefore, an in-depth understanding of the pathogenesis of spinal cord compression injury is required to help develop effective clinical interventions. In the present study, we propose a new method of quantitative 3D micro-CT to observe microvascular events in a chronic spinal cord compression rat model. A total of 36 rats were divided into two groups: sham control group (n = 12) and compressive spinal cord injury group (n = 24). Rats were scarified at four weeks after surgery. In each group, CD34 micro-vessel immunohistochemical staining was performed in half of the animals, while micro-CT scanning was performed in the other half. Microvessel density (MVD) was measured after immunohistochemical staining, while the vascular index (VI) was measured in 3D micro-CT. In comparison with sham control, abnormal somatosensory evoked potentials (SEP) can be seen in all 24 cases of the compression group, and VI shows the amount of microvessels reduced consistently and significantly (p < 0.01). A significant correlation is also found between MVD and VI (r = 0.95, p < 0.01). These data suggest that quantitative 3D micro-CT is a sensitive and promising tool for investigating microvascular changes during chronic compressive spinal cord injury.

Aim: To establish a Chinese esophageal squamous cell carcinoma (ESCC) cell line with high bone metastasis potency using (99m)Tc-methylene diphosphonate ((99m)Tc-MDP) micro-pinhole scintigraphy, X ray and micro-positron emission tomography/computed tomography (PET/CT) for exploring the mechanism of occurrence and development in esophageal cancer., Methods: The cells came from a BALB/c nu/nu immunodeficient mouse, and oncogenic tumor tissue was from a surgical specimen from a 61-year-old male patient with ESCC. The cell growth curve was mapped and analysis of chromosome karyotype was performed. Approximately 1 × 10⁶ oncogenic cells were injected into the left cardiac ventricle of immunodeficient mice. The bone metastatic lesions of tumor-bearing mice were detected by (99m)Tc-MDP scintigraphy, micro-PET/CT and X-ray, and were resected from the mice under deep anesthesia. The bone metastatic cells in the lesions were used for culture and for repeated intracardiac inoculation. This in vivo/in vitro experimental metastasis study was repeated for four cycles. All of the suspicious bone sites were confirmed by pathology. Real-time polymerase chain reaction was used to compare the gene expression in the parental cells and in the bone metastatic clone., Results: The surgical specimen was implanted subcutaneously in immunodeficient mice and the tumorigenesis rate was 100%. First-passage oncogenic cells were named CEK-Sq-1. The chromosome karyotype analysis of the cell line was hypotriploid. The bone metastasis rate went from 20% with the first-passage oncogenic cells via intracardiac inoculation to 90% after four cycles. The established bone metastasis clone named CEK-Sq-1BM had a high potential to metastasize in bone, including mandible, humerus, thoracic and lumbar vertebrae, scapula and femur. The bone metastasis lesions were successfully detected by micro-pinhole bone scintigraphy, micro-PET/CT, and X-ray. The sensitivity, specificity and accuracy of the micro-pinhole scintigraphy, X-ray, and micro-PET/CT imaging examinations were: 89.66%/32%/80%, 88.2%/100%/89.2%, and 88.75%/77.5%/87.5%, respectively. Some gene expression difference was found between parental and bone metastasis cells., Conclusion: This newly established Chinese ESCC cell line and animal model may provide a useful tool for the study of the pathogenesis and development of esophageal carcinoma.

Surface wrinkling of mucosae is crucial for the biological functions of many living tissues. In this paper, we investigate the instability of a cylindrical tube consisting of a mucosal layer and a submucosal layer. Our attention is focused on the effects of internal pressure and surface tension on the critical condition and mode number of surface wrinkling induced by tissue growth. It is found that the internal pressure plays a stabilizing role but basically has no effect on the critical mode number. Surface tension also stabilizes the system and reduces the critical mode number of surface patterns. Besides, the thinner the mucosal layer, the more significant the effect of surface tension. This work may help gain insights into the surface wrinkling and morphological evolution of such tubular organs as airways and esophagi., (Copyright © 2013 Elsevier Ltd. All rights reserved.)

Objective: To evaluate the effect of expansion speed on chronic compressive spinal cord injury in the rat., Methods: Thirty-six Sprague-Dawley rats were divided into four groups: a control group, a group receiving compressor in the C5-C6 epidural space with instant compression (group 1), and two other groups receiving water-absorbing polyurethane polymer sheets with two expansion speeds, which reached maximum volume in 2 h (group 2: fast expansion) or 24 h (group 3: slow expansion). A C6 laminectomy was performed in the control group. Neurological function, MRI, large motoneuron number in the ventral horn, and myelin staining intensity in the posterior funiculus were evaluated., Results: In the instant compression group, compression was confirmed on T2-weighted images by a hypointense signal change in the intramedulla. In the gradual compressive injury groups, large motoneuron number (p<0.001), but not myelin staining intensity, was significantly decreased in both the fast and slow expansion groups compared with the instant compression group. However, there was no difference in Basso Beattie Bresnahan score, cord distortion in T2-weighted image, large motoneuron numbers, or myelin staining between the fast and slow expansion groups., Conclusion: Instant spinal cord compression caused acute injury. Gradual expansion compression induced reliable pathology and MRI characteristics consistent with chronic compressive spinal cord injury. The speed of expansion is not a significant problem for establishing a reliable model if the chronic compression is induced by gradual expansion., (Copyright © 2013 Elsevier Ireland Ltd. All rights reserved.)

Objective: The aim of this study was to evaluate the prevalence and spectrum of Nkx2.5 mutations associated with idiopathic atrial fibrillation (AF)., Methods: A cohort of 136 unrelated patients with idiopathic atrial fibrillation and 200 unrelated, ethnically matched healthy controls were enrolled. The coding exons and splice junctions of the Nkx2.5 gene were sequenced in 136 atrial fibrillation patients, and the available relatives of mutation carriers and 200 controls were subsequently genotyped for the identified mutations. The functional characteristics of the mutated Nkx2.5 gene were analyzed using a dual-luciferase reporter assay system., Results: Two novel heterozygous Nkx2.5 mutations (p.N19D and p.F186S) were identified in 2 of the 136 unrelated atrial fibrillation cases, with a mutational prevalence of approximately 1.47%. These missense mutations co-segregated with atrial fibrillation in the families and were absent in the 400 control chromosomes. Notably, 2 mutation carriers also had congenital atrial septal defects and atrioventricular block. Multiple alignments of the Nkx2.5 protein sequences across various species revealed that the altered amino acids were completely conserved evolutionarily. Functional analysis demonstrated that the mutant Nkx2.5 proteins were associated with significantly reduced transcriptional activity compared to their wild-type counterpart., Conclusion: These findings associate the Nkx2.5 loss-of-function mutation with atrial fibrillation and atrioventricular block and provide novel insights into the molecular mechanism involved in the pathogenesis of atrial fibrillation. These results also have potential implications for early prophylaxis and allele-specific therapy of this common arrhythmia.

Statistically based experimental designs were applied to optimize the fermentation medium and cultural conditions for the maximization of neutral protease using three agroindustrial residues (cassava pulp, soybean meal, and wheat bran) and Bacillus subtilis DES-59. The Plackett-Burman design was used to evaluate the effects of variables such as the concentration of substrates, initial pH, shaker's rotating speed, temperature, inoculum size, and incubation time. Among the eight parameters, three significant variables (cassava pulp, soybean meal, and inoculum size) were selected for the optimization study, in which a central composite design was used to optimize the concentrations of cassava pulp and soybean meal and inoculum size and investigate the interactive effects of the three variables. The optimal parameters obtained from response surface methodology are 37.78 g/L of cassava pulp, 15 g/L of soybean meal, and 6.5% (v/v) of inoculum size, respectively, resulting in a maximum neutral protease activity of 4107 ± 122 U/mL., (© 2013 International Union of Biochemistry and Molecular Biology, Inc.)

The marine diatom Phaeodactylum tricornutum, a widely used forage species, has a storage lipid content of up to 30% dry cell weight. To explore the mechanism behind the high storage lipid accumulation in this diatom, acetyl-CoA carboxylase (ACCase), which catalyzes the first committed step of the fatty acid biosynthetic pathway, was characterized in this study. A homogeneous type of ACCase (PtACC) was identified from P. tricornutum by homology searches. The first exon of the ACCase gene (PtACC-1) was cloned. PtACC-1 was fused with a Myc epitope tag and cloned into plasmid pMD18 driven by the LacZ promoter and expressed in Escherichia coli. The expression of the PtACC-1-Myc protein was verified by Western blot. The neutral lipid content in transformed E. coli increased substantially by twofold as determined by Nile red fluorescent dye staining. Concomitantly, ACCase activity increased by 1.72-fold. The fatty acid composition, analyzed by GC-MS, demonstrated a significant difference in the ratio of saturated fatty acids and monounsaturated fatty acids (MUFAs). MUFAs of PtACC-1 expressing cells increased by 13%. This study represents the first characterization of the key domains of ACCase from a diatom and demonstrates high neutral lipid accumulation in E. coli expressing PtACC-1, providing an additional genetic resource with the potential for biodiesel development., (© 2013 International Union of Biochemistry and Molecular Biology, Inc.)

Cervical spondylotic myelopathy (CSM) is one of the most common spinal cord disorders affecting the elderly. Yet the exact pathophysiology of CSM remains unclear. Vascular response to initial mechanical compression and associated ischemia may involve in secondary pathophysiology. Chronic compressive lesions to cervical cord resulting in lack of perfusion have established considerable evidences to support ischemia as an important pathogenesis both in patients and animal models, a similarity as that of acute spinal cord injury (SCI). In hypoxic condition following SCI, the up-regulation of vascular endothelial growth factor (VEGF), is consistent with increasing hypoxia induced factor-1α (HIF-1α) in acute periods. HIF-1α/VEGF signaling pathway is thought to play a dual role following SCI. In one hand, VEGF was demonstrated to be correlated with angiogenesis (protecting vascular endothelial cells, increasing blood vessel density and improving regional blood flow), neurogenesis (antiapoptotic, neurotrophic, attenuate axonal degradation), and locomotor ability improvement. In other hand, some studies revealed that VEGF have limited therapeutic effect, even exacerbate the secondary damage following SCI. VEGF administrations in acute or subacute periods result in elevation of blood-spinal cord barrier (BSCB) permeability even last for chronic course. BSCB permeability elevation initiates a secondary cascade of events involving excitotoxicity, infiltration of leukocytes and tissue edema. With comprehensive understanding of temporal and spatial of HIF-1α/VEGF signaling pathway, development of therapeutic strategies to promote new vessel growth while minimize the deleterious effects of VEGF-induced microvascular permeability, and thereby improve neurologic function, seems to be feasible and promising., (Copyright © 2012 Elsevier Ltd. All rights reserved.)

Genetic transformation is useful for basic research and applied biotechnology. However, genetic transformation of microalgae is usually quite difficult due to the technical limitations of existing methods. We cloned the promoter and terminator of the nitrate reductase gene from the microalga Phaeodactylum tricornutum and used them for optimization of a transformation system of the microalga Chlorella vulgaris. This species has been used for food production and is a promising candidate as a bioreactor for large-scale production of value-added proteins. A construct was made containing the CAT (chloramphenicol acetyltransferase) reporter gene driven by the nitrate reductase promoter. This construct was transferred into the C. vulgaris genome by electroporation. Expression of CAT in transgenic Chlorella conferred resistance to the antibiotic chloramphenicol and enabled growth in selective media. Overall efficiency for the transformation was estimated to be approximately 0.03%, which is relatively high compared with other available Chlorella transformation systems. Expression of CAT was induced in the presence of nitrate and inhibited in the presence of ammonium as a sole nitrogen source. This study presented an inducible recombinant gene expression system, also providing more gene regulation elements with potential for biotechnological applications.

Objective: To investigate the drug distribution in tissues of cervical lymph node metastasis mice model after submucosa adjacent cancer injection of pingyangmycin-activated carbon nanoparticles (PYM-CH-NP) and evaluate the lymph targeting effect of PYM-CH-NP., Methods: Pingyangmycin (PYM) was radiolabeled with 125I by modified the chloramine T method. Cervical lymph node metastasis mice model was established by buccal submucosa inoculation of a high lymph metastasis cell line U14 cancer cell. 360 mice models burdened with cervical lymph metastasis were randomly divided into 3 groups. PYM group was treated with PYM water solution, PYM-CH-NP group was treated with PYM-CH-NP. Negative control group was injected with activated carbon nanoparticles. PYM-CH-NP and pingyangmycin water solution were injected in pericancer submucosa of the mice respectively. The radioactivity of drug in blood, heart, liver, spleen, lung, kidney and cervical lymph node were measured after 0.5, 1, 4, 8, 12, 24, 48, 72, 96, 120, 144, 168 h administration. The radioactivity of each samples per unit weight were calculated. The selectivity index (SI) and targeting index (TI) of drug were calculated., Results: The radioactivity of drug in cervical lymph node of PYM-CH-NP group was much higher than PYM group in each time point (P < 0.001), whereas the blood, heart, liver, spleen, lung and kidney uptake of pingyangmycin was greatly decreased in PYM-CH-NP group after 4 h administration (P < 0.001). The SI value of PYM group at each time point was less than 1. While the minimum SI and TI value of PYM-CH-NP was 1.793 and 1.562, the maximum value reached to 68.126 and 14.623 after 72 h administration., Conclusion: PYM-CH-NP can increase drug dosage in metastasized cervical lymph nodes, and decrease drug dosage of other organs. So better therapeutic outcome and little adverse reaction may be achieved for lymph node metastasis.

Objective: To study the allocation of embedded teeth in jaws using the three-dimensional reconstruction technique of 64-slices spiral CT., Methods: 27 cases were examined by helical scanning of axial view volume scan CT. The exact localization of the embedded teeth in jaws was acquired by using volume rendering (VR), maximum intensity projection (MIP), multiplanar reformation (MPR) and curve plane reconstruction (CPR)., Results: The localization, morphous, size, erupted orientation and relationship between surrounding tissues of the 41 embedded teeth in 27 patients were displayed by effectually using the images of VR, MIP, MPR and CPR. The election of orthodontic treatment or surgical intervention was decided by using 64-slices spiral CT., Conclusion: The exact data and objective evidence of the treatment plan could be provided by 64-slices spiral CT which will have important clinical application.

Objective: To investigate the effect of the traditional Chinese medicine Matrine on cell cycle and human telomerase reverse transcriptase (hTERT) of human ACC-M cell lines., Methods: Different concentrations of Matrine were used in the medium of ACC-M cells. Change of cell cycle were detected by flow cytometry after ACC-M cell were cultivated with different concentrations Matrine (0.25, 0.50, 0.75, 1.00 mg x mL(-1)). Expression of hTERT was investigated by reverse transcription-polymerase chain reaction (RT-PCR) and indirect immunofluorescene and flow cytometry quantitative analysis., Results: Matrine caused obviously the GdG1 phase block and inhibited proliferation of ACC-M cells. At same time, this effect was positive correlation to Matrine concentration and treat time. Matrine can inhibit the expression of hTERT mRNA and protein., Conclusion: Matrine can obviously inhibit cell cycle and down-regulate expression of hTERT. Inhibition of cell cycle is possible correlation with down-regulation expression of hTERT.

This paper reported a 16-year-old girl suffering from painless swelling in the right submandibular region which was diagnosed as ranula, but during the surgery, it was seen that the cyst was closely related to the submandibular gland and it was proved by pathological diagnosis as submandibular cyst. Submandibular cyst and ranula should be distinguished clinically, in order to avoid misdiagnosis and mistreatment.

Simulations of gastrointestinal digestion were used to try to identify the nature of the complexes between antinutritional factors and iron and zinc in faba bean and legume fractions. In digestible residue of raw faba bean flour, simultaneous action of cellulase and phytases made it possible to release about 28% units more iron than that released with the treatment without enzymes. About 49.8% of iron in raw faba bean flour was solubilized after in vitro digestion and simultaneous action of cellulase and phytase. In the hull fraction, the action of phytases and the simultaneous action of cellulase and phytase allowed about 7 and 35% units of additional zinc to be solubilized, respectively. Single enzymatic degradation of phytates from dehulled faba bean allowed solubilization from 65 to 93% of zinc, depending upon the treatment. In dehulled faba bean, iron was chelated by phytates and by fibers, whereas zinc was almost exclusively chelated by phytates. In the hull of faba bean, a high proportion of iron was chelated by iron-tannins, while the rest of iron as well as the majority of zinc were chelated in complexes between phytates and fibers.

Purpose: To investigate the effect and mechanism of matrine on adhesion and invasion of human adenoid cystic carcinoma ACC-M cells in vitro., Methods: MTT assay was used to examine the effect of matrine on proliferation of ACC-M cells treated after 1 day to 3 days. Byden chamber assay was performed to detect the effect of matrine on invasion capacity of the cells; Effect on adhesion potential of ACC-M cells was tested by cell matrigel adhesion assay. Mechanism of inhibitory adhesion and invasion was investigated by expression of E-cad protein. The results was analyzed by SPSS11.0 software package., Results: Matrine could inhibit the proliferation of ACC-M cell lines with apparent dose-dependent and time-dependent effect. Matrine significantly inhibited adhesion and invasion capacity of ACC-M cell lines in vitro with concentration increasing. The expression level of E-cad was significantly increased compared with the untreated group in ACC-M cell lines., Conclusions: Matrine can inhibit the adhesion and invasion capacity of ACC-M cell lines in vitro. Mechanism of the inhibition effect may be related to over expression of E-cad protein.

Purpose: The purpose of this study was to explore the diagnosis and operative methods for traumatic temporomandibular joint ankylosis., Methods: Fourteen patients (8 females, 6 males; aged from 7 to 23 years, median age 17.5 years) with traumatic TMJ ankylosis of 1 to 15 years' duration, with a maximal mouth opening(MMO) from 0 to 1.0cm(average:0.46cm) preoperatively were included in this study. They were further divided into two types(I and II) based on false articulation medial to the analysis as shown by coronal computed tomography (CT) coronoid scan. 7 TMJs were treated with medial arthroplasty(MA), in which the lateral fusional bone was removed and the medial false articulation was retained for type I, 6 TMJs were treated with natural positional arthroplasty(NPA) in which the bony fusion between the condyle and glenoid fossa was separated and shaved, the disc was retained and repositioned for type II. Follow-up was carried out and the surgical effect was assessed based on clinical and imaging findings. Student's t test was conducted for comparison with SPSS15.0 software package., Results: There was 9 TMJs from 7 patients for type I and 5 TMJs from 5 patients for type II as shown by CT. The mean postoperative MMO was 3.12cm, and the condylar shape was smooth on CT scan. There was significant difference between preoperative and postoperative MMO, P<0.01. There was no significant difference between MA and NPA, P>0.05., Conclusions: It is concluded that type I is more common in traumatic TMJ ankylosis than type II. The surgical effect of MA and NPA is good, with less injury, and is worthy of wide application.

Objective: To study the molecular genetic etiology of a Chinese pedigree with basal cell nevus syndrome., Methods: The proband and his affected mother and a unaffected individual in the pedigree were chosen and peripheral blood was collected from them for DNA. Direct sequencing was performed to detect the mutations of PTCH gene. In order to further confirm the results of sequence analysis, all available family members were analyzed with genetic linkage analysis using 3 highly polymorphic microsatellite DNA markers in the region of 9q22.3-q31., Results: No mutations of PTCH gene was detected in the proband's mother, one synonymous mutation was detected in the proband. Linkage analysis showed that the Lod scores of the 3 markers were: D9S283, Z = -2.11 (theta = 0.00); D9S1690, Z = -2.95 (theta = 0.00); D9S1677, Z = -5.94 (theta = 0.00)., Conclusions: In this pedigree, mutation of PTCH gene is not related to the underlying pathogenesis of the syndrome.

Background: Screening libraries against a molecular target in vitro are idealized models that cannot reflect the real state in vivo where biomolecules coexist and interact. C-terminal amide tripeptides labelled with Technetium-99m can provide a unique noninvasive approach to trace a large number of compounds in vivo., Methods: The C-terminal amide tripeptide libraries were synthesized on Rink Amide-MBHA resin using iterative and pooling protocol. Technetium (V) oxo core [TcO(3+)] was bound to each tripeptide via 4 deprotonated nitrogen atoms to form a library of 8000 (99m)Tc tripeptoid complexes. The radiocombinatorial screening (RCS) in vivo was carried out on SD rats and A549 tumour bearing mice., Results: Signals of tissue distribution and metabolism of libraries were recorded by counting or imaging and tissue targeting leads identified by both random and directed RCS. Among them, (99m)Tc RPA, (99m)Tc VIG and (99m)Tc RES had specific tissue targeting in kidney, liver and tumour respectively. The percent injected dose per gram tissue of (99m)Tc labelled leads in their target tissue was highly structure dependent. Because the nontarget tissue binding and the metabolism of (99m)Tc tripeptoid sublibraries were simultaneously monitored successfully by RCS, the interference of background activity was limited to the lowest level. Optimization of renal function agent from the labelled libraries was carried out by directed screening. (99m)Tc DSG was finally identified the most promising agent for renal function studies., Conclusions: RCS in vivo is a powerful tool for the discovery of tissue targeting drugs. The potential screening bias is probably the major limitation of labelled libraries.

Dynamic analysis of viral nucleic acids in host cells is important for understanding virus-host interaction. By labeling endogenous RNA with molecular beacon, we have realized the direct visualization of viral nucleic acids in living host cells and have studied the dynamic behavior of poliovirus plus-strand RNA. Poliovirus plus-strand RNA was observed to display different distribution patterns in living Vero cells at different post-infection time points. Real-time imaging suggested that the translocation of poliovirus plus-strand RNA is a characteristic rearrangement process requiring intact microtubule network of host cells. Confocal-FRAP measurements showed that 49.4 +/- 3.2% of the poliovirus plus-strand RNA molecules diffused freely (with a D-value of 9.6 +/- 1.6 x 10(-10) cm2/s) within their distribution region, while the remaining (50.5 +/- 2.9%) were almost immobile and moved very slowly only with change of the RNA distribution region. Under the electron microscope, it was found that virus-induced membrane rearrangement is microtubule-associated in poliovirus-infected Vero cells. These results reveal an entrapment and diffusion mechanism for the movement of poliovirus plus-strand RNA in living mammalian cells, and demonstrate that the mechanism is mainly associated with microtubules and virus-induced membrane structures.

A new approach, short-oligonucleotide-ligation assay on DNA chip (SOLAC), is developed to detect mutations in rifampin-resistant Mycobacterium tuberculosis. The method needs only four common probes to detect 15 mutational variants of the rpoB gene within 12 h. Fifty-five rifampin-resistant M. tuberculosis isolates were analyzed, resulting in 87.3% accuracy and 83.6% concordance relative to DNA sequencing.

Homogeneity is proposed for evaluation of the quality of analytical biodevices, such as biosensors and biochips. As a demonstration, glucose oxidase (GOx) was modified at its C-terminal with a linker peptide with a cysteine residue at the end. The fusion structure (GOx-linker-cysteine) enables the enzyme to immobilize on gold surfaces with a Cys-S-Au bond or to immobilize on a silanized glass surface via disulfide chemistry. With this fusion structure, the enzyme can be anchored onto the substrate with well-controlled orientation, thus forming a homogeneous biological layer on biodevices. The linker peptide between GOx and the cysteine acts as a spacer to reduce the steric hindrance caused by the bulky body of the enzyme. Biochemistry experiments showed that this genetically modified glucose oxidase (shortened to GOxm) retained most of its catalytic characteristics, with K(m) and K(cat) similar to those of the wild-type GOx. Electrochemistry experiments showed that GOxm-modified electrode gave higher and more stable current responses than the electrode modified with GOx which has no free -SH on its surface. The coefficients of variation (used for evaluation of the interchangeability of the enzyme device from the same batch preparation) were 9.5% for the GOxm gold electrode and 20.0% for the GOx gold electrode and the GOxm oxygen electrode. The relative errors (used for evaluation of the precision of the individual enzyme device) were 2.9% for the GOxm gold electrode, 12.0% for the GOx gold electrode, and 11.2% for the GOxm oxygen electrode. Atomic force microscopy images revealed that GOxm formed a self-assembled monolayer in a hexagonal-like lattice packing arrangement on the gold surface, while GOx formed multilayer assembling or aggregated particles. The homogeneity of the protein chips, the GOxm array that was prepared through -S-S- formation, and the GOx array that was prepared through nonspecific adsorption was evaluated. The coefficients of variation, calculated with the signal level of all dots, were 5.4% for the GOxm array and 81.8% for the GOx array. All experimental results pointed to the fact that the homogeneity of the analytical biodevices could be considerably improved by using the proposed method.

An accurate density-functional method is used to study systematically half-metallic ferromagnetism and stability of zincblende phases of 3d-transition-metal chalcogenides. The zincblende CrTe, CrSe, and VTe phases are found to be excellent half-metallic ferromagnets with large half-metallic gaps (up to 0.88 eV). They are mechanically stable and approximately 0.31-0.53 eV per formula unit higher in total energy than the corresponding nickel-arsenide ground-state phases, and therefore would be grown epitaxially in the form of films and layers thick enough for spintronic applications.

Objective: To elucidate the molecular mechanism of X-linked adrenoleukodystrophy(ALD) in Chinese., Methods: Polymerase chain reaction in exon 1, exon 5 and their flanking sequences and direct DNA sequencing of ALD gene were performed in four patients, their mothers and twenty normal individuals as controls., Results: A splice mutation was identified in the interface of exon 5 and intron 5 (1875 G-->A). This splice mutation in 5' end of intron 5 might lead to abnormal splice in exon 5 and exon 6 and bring about unstable and abnormal ALD protein; the lignoceryl CoA ligase could not transport very long chain fatty acids (VLCFA) into peroxisome and could not function normally; consequently, defective beta-oxidation of VLCFA in peroxisome could result in an accumulation of VLCFAS in the central nervous system, adrenal gland and blood., Conclusion: The splice mutation in 5' end of intron 5 leading to abnormal splice in exon 5 and exon 6 appears to be one of the causes of X-linked recessive adrenoleukodystrophy.

Glucose oxidase (GOD) was genetically modified by adding a poly-lysine chain at the C-terminal with a peptide linker inserted between the enzyme and poly-lysine chain. The poly-lysine chain was added in order to anchor more electron transfer mediator, ferrocenecarboxylic acid, to GOD for the purpose of improving sensitivity and stability of glucose biosensors. The modified GOD had similar K(m) and K(cat) to those of the wild type enzyme. After interacted with the electron transfer mediator, the modified enzyme retained 90.01% of its native activity, while the commercial GOD and the wild type GOD (Aspergillus niger) retained only 22.43 and 22.17%, respectively. Screen-printed electrodes coated with the modified GOD, wild type yeast-derived GOD or the commercial GOD were tested in glucose solution of different concentrations. Experimental results showed that the biosensor based on the modified GOD gave the largest signal among the three. In addition, the linear range of the biosensor prepared by the modified GOD could extend to 45 mM, while they were about 20 mM for the biosensors based on the wild type yeast-derived enzyme and the commercial enzyme.