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To investigate the effects of hypertension on the changes in gap junctions between vascular smooth muscle cells (VSMCs) in the mesenteric artery (MA) of spontaneously hypertensive rats (SHRs). Whole-cell patch clamp, pressure myography, real-time quantitative reverse transcription PCR (qRT-PCR), western blot analysis and transmission electron microscopy were used to examine the differences in expression and function of the gap junction between MA VSMCs of SHR and control normotensive Wistar-Kyoto (WKY) rats. (1) Whole-cell patch clamp measurements showed that the membrane capacitance and conductance of in-situ MA VSMCs of SHR were significantly greater than those of WKY rats (P<0.05), suggesting enhanced gap junction coupling between MA VSMCs of SHR. (2) The administration of phenylephrine (PE) and KCl (an endothelium-independent vasoconstrictor) initiated more pronounced vasoconstriction in SHR versus WKY rats (P<0.05). Furthermore, 2-APB (a gap junction inhibitor) attenuated PE- and KCl-induced vasoconstriction, and the inhibitory effects of 2-APB were significantly greater in SHR (P<0.05). (3) The expression of connexin 45 (Cx45) mRNA and protein in the MA was greater in SHR versus WKY rats (P<0.05). The level of phosphorylated Cx43 was significantly higher in SHR versus WKY rats (P<0.05), although the expression of total Cx43 mRNA and protein in the MA was equivalent between SHR and WKY rats. Electron microscopy revealed that the gap junctions were significantly larger in SHR versus WKY rats. Increases in the expression of Cx45 and phosphorylation of Cx43 may contribute to the enhancement of communication across gap junctions between MA VSMCs of SHR, which may increase the contractile response to agonists.

Background: Endoscopic retrograde cholangiopancreatography with fluoroscopy guidance is a well-established technique for providing biliary drainage in patients with biliary obstructions. However, fluoroscopic facilities may not always be available and fluoroscopy carries a risk of radiation exposure., Aim: We retrospectively compared the procedure success rate and efficacy of ultrasound-guided endoscopic biliary drainage (UG-EBD) and fluoroscopy-guided endoscopic biliary drainage (FG-EBD) in patients with biliary obstructions., Methods: Patients who had received either UG-EBD or FG-EBD were included in the study. Main outcome measurements included the procedure success rate, procedure time, and clinical response., Results: A total of 125 patients who had undergone UG-EBD (n = 63) and FG-EBD (n = 62) were identified. The total procedure success rate was 93.7 % in the UG-EBD group and 96.8 % in the FG-EBD group without statistical difference. Also, no significant difference was found in the procedure success rate of lower or upper/middle obstructions of the common bile duct (CBD) between the 2 groups. The mean procedure time was not different between the 2 groups [UG-EBD group 24.54 (9.52) min vs. FG-EBD group 21.74 (8.77) min, p = 0.09]. There were no differences in the normalization of clinical and laboratory parameters and immediate complication between the 2 groups., Conclusions: Endoscopic biliary drainage (EBD) under US-guidance and under fluoroscopy guidance is equally effective and safe for patients with lower or upper/middle obstructions of the CBD. The UG-EBD technique is especially suitable for special patients, such as critically ill patients, pregnant woman, etc.

Pollution Status and characteristics of PBDEs in offices were investigated in Hangzhou. As a result, the total concentration of PBDEs was 40.66-141.00 pg x m(-3), and the mean concentration was 93.22 pg x m(-3), being 1.87 and 5.01 times as high as those in homes and outdoor. In particle and gas phases, BDE-47 and BDE-99 were the most abundant congeners, which accounted for 33.29% and 31.99% of total PBDEs, respectively. Concentration of PBDEs in gas phase was 1.34 times as high as that in particle phase. BDE-28, BDE-47 and BDE-99 mainly existed in the gas phase, while BDE-153 and BDE-183 mainly existed in the particle phase.

Objective: This study investigated the effectiveness and safety of topical application of diosmectite (DS) and basic fibroblast growth factor (bFGF) paste in the treatment of minor recurrent aphthous stomatitis., Study Design: Four pastes, containing bFGF and DS, DS alone, bFGF alone, and vehicle only, were consecutively applied 4 times per day for 5 days. Pain levels and the amount of ulcer amelioration were evaluated., Results: A total of 129 participants completed the study. DS+bFGF paste released bFGF continuously and significantly lowered ulcer pain scores (P < .05 for days 3, 4, 5, and 6) compared with the other pastes. The ulcer size was significantly reduced (P < .05 for days 2, 4, and 6) in the DS+bFGF paste group compared with the other groups. No obvious adverse drug effects were observed., Conclusions: DS+bFGF paste was effective in alleviating ulcer pain and promoting healing without significant side effects in the treatment of minor recurrent aphthous stomatitis., (Copyright © 2013 Elsevier Inc. All rights reserved.)

Background/aims: Endotoxin (lipopolysaccharide, LPS)-induced acute liver injury was attenuated by endotoxin tolerance (ET), which is characterized by phosphatidylinositol 3-kinase pathway/Akt signaling. Glycogen synthase kinase 3 (GSK-3) acts downstream of phosphatidylinositol 3-kinase pathway/Akt and GSK-3 inhibitor protects against organic injury. This study evaluates the hypothesis that ET attenuated LPS-induced liver injury through inhibiting GSK-3 functional activity and downstream signaling., Methods: Sprague-Dawley rats with or without low-dose LPS pretreatment were challenged with or without large dose of LPS and subsequently received studies. Serum tumor necrosis factor-alpha, interleukin-10, alanine aminotransferase, lactate dehydrogenase, and total bilirubin levels were analyzed, morphology of liver tissue was performed, glycogen content, myeloperoxidase content, phagocytosis activity of Kupffer cells, and the expression and inhibitory phosphorylation as well as kinase activity of GSK-3 were examined. Survival after LPS administration was also determined., Results: LPS induced significant increases of serum TNF-α, alanine aminotransferase, lactate dehydrogenase, and total bilirubin (P < 0.05), which were companied by obvious alterations in liver: the injury of liver tissue, the decrease of glycogen, the infiltration of neutrophils, and the enhancement of phagocytosis of Kupffer cells (P < 0.05). LPS pretreatment significantly attenuated these alterations, promoted the inhibitory phosphorylation of GSK-3 and inhibited its kinase activity, and improved the survival rate (P < 0.05)., Conclusions: ET attenuated LPS-induced acute liver injury through inhibiting GSK-3 functional activity and its downstream signaling., (Copyright © 2013 Elsevier Inc. All rights reserved.)

The genus Amphibacillus was established in 1990, and seven additional species were described in the past two decades. Amphibacillus jilinensis Y1(T) is a facultatively anaerobic and alkaliphilic bacterium isolated from a soda lake in China. Here we describe the structural and genetic features of the draft genome about the type strain Y1(T) (3,831,075 bp, with a G+C content of 37.27%). This is the first genome report of the Amphibacillus genus.

Objective: This study compared Wistar rat with spontaneously hypertensive rat (SHR) on the electrophysiology and coupling force of the smooth muscle cells in the cerebral arteriolar segments and observe the influence of 18beta-glycyrrhetinic acid(18beta-GA) on the gap junctions between the arterial smooth muscle cells., Methods: The outer layer's connective tissue of the cerebral arteriolar segments was removed. Whole-cell patch clamp recordings were used to observe the 18beta-GA's impaction on the arteriolar segment membrane's input capacitance (C(input)), input conductance (G(input)) and input resistance (R(input)) of the smooth muscle cells., Results: (1) The C(input) and G(input) of the SHR arteriolar segment smooth muscle cells was much higher than the Wistar rats, there was significant difference (P < 0.05). (2) 18beta-GA concentration-dependently reduced C(input) and G(input) (or increase R(input)) on smooth muscle cells in arteriolar segment. IC50 of 18beta-GA suppression's G(input) of the Wistar rat and SHR were 1.7 and 2.0 micromol/L respectively, there was not significant difference (P > 0.05). After application of 18beta-GA concentration > or = 100 micrmol/L, the C(input), G(input) and R(input) of the single smooth muscle cells was very close., Conclusion: Gap junctional coupling is enhanced in the SHR cerebral arterial smooth muscle cells. 18beta-GA concentration-dependent inhibits Wistar rat's and SHR cerebral arteriolar gap junctions between arterial smooth muscle cells. The inhibitory potency is similar between the two different rats. When 18beta-GA concentration is > or = 100 micromol/L, it can completely block gap junctions between arteriolar smooth muscle cells.

Long noncoding RNAs (lncRNAs) are pervasively transcribed and have a critical role in genome regulation. Alterations in the expression of several lncRNAs have been observed in some types of cancers; however, the contributions of lncRNAs to osteosarcoma remain unknown. Here, we describe the expression profile of lncRNAs in osteosarcomas compared with paired adjacent noncancerous tissue using microarray analysis. In our study, 25,733 lncRNAs were expressed in osteosarcoma; 403 lncRNAs were consistently over-regulated and 798 lncRNAs were consistently under-regulated in all samples analyzed (⩾2.0-fold, p<0.05). Quantitative real-time polymerase chain reaction (PCR) was used to validate six over-regulated and four under-regulated lncRNAs. Bioinformatic analysis (gene ontology analysis, pathway analysis and network analysis) was used for further research. Pathway analysis indicated that 32 pathways corresponded to under-regulated transcripts and that 34 pathways corresponded to over-regulated transcripts (p-value cut-off is 0.05). Our results are the first to reveal differentially expressed lncRNAs in osteosarcoma and suggest that lncRNAs may be novel candidate biomarkers for the diagnosis of osteosarcoma and potential targets for therapy., (Copyright © 2013 Elsevier Inc. All rights reserved.)

Spiral ganglion neurons (SGNs) are poorly regenerated in the mammalian inner ear. Because of this, stem cell transplantation has been used to replace injured SGNs, and several studies have addressed this approach. However, the difficulty of delivering stem cells into the cochlea and encouraging their migration to Rosenthal's canal (RC), where the SGNs are located, severely restricts this therapeutic strategy. In this study, we attempted to establish a new stem cell transplantation route into the cochlea via the cochlear lateral wall (CLW). First, we tested the precision of this route by injecting Fluorogold into the CLW and next assessed its safety by mock surgeries. Then, using a degenerated SGN animal model, we transplanted neural stem cells (NSCs), derived from the olfactory bulb of C57BL/6-green fluorescent protein (GFP) mice, via the CLW route and examined the cells' distribution in the cochlea. We found the CLW transplantation route is precise and safe. In addition, NSCs migrated into RC with a high efficiency and differentiated into neurons in a degenerated SGN rat model after the CLW transplantation. This result revealed that the basilar membrane (BM) may have crevices permitting the migration of NSCs. The result of this study demonstrates a novel route for cell transplantation to the inner ear, which is important for the replacement of degenerated SGNs and may contribute to the treatment of sensorineural hearing loss., (Copyright © 2013 Elsevier B.V. All rights reserved.)

Objective: The aim of this study was to investigate the effectiveness of triamcinolone acetonide (TA) and salvianolic acid B (SA-B) intralesional combined injection in the treatment of oral submucous fibrosis (OSF)., Material and Methods: A randomized clinical trial was performed. TA, SA-B, and TA combined with SA-B were consecutively applied intralesionally weekly for 20 weeks. Mouth opening and burning sensation improvement as determined by a 100-mm visual analog scale were evaluated at weeks 10, 20, and 44., Results: Forty-two subjects fulfilled the study without obvious adverse reactions. The net gain in mouth opening was 2.00 ± 1.21 mm in the TA group, 3.48 ± 2.23 mm in the SA-B group, and 5.50 ± 1.80 mm in the TA + SA-B group at week 44. The burning sensation improved by 3.05 ± 0.76 in the TA group, 4.96 ± 0.97 in the SA-B group, and 6.11 ± 0.93 in the TA + SA-B group by the end of the study., Conclusions: TA + SA-B intralesional injections improved mouth open and burning sensation in these OSF patients., (Copyright © 2013 Elsevier Inc. All rights reserved.)

Neural stem cell (NSC) transplantation into the cochlea is widely used for the treatment of spiral ganglion neuron (SGN) degenerative disease and injury in the animal models, but the migration of the transplanted NSCs to the injury region is difficult and the mechanism is still unclear. In this study, we aimed to validate whether the SGN-degenerated cochlear microenvironment plays a role in the NSC migration and investigated whether stromal cell-derived factor-1 (SDF-1) was involved in the NSCs migration. Using a rat SGN degeneration model, we demonstrated that the transplanted NSCs are more likely to migrate to the injury region during the early post-injury (EPI) than the late post-injury (LPI) stage and the control cochlea. We found that the expressions of SDF-1 increased transiently after SGN degeneration. Additionally, we showed that the NSCs express CXCR4, a receptor for SDF-1. We observed that the region to which the transplanted NSC localized coincides with the region where the SDF-1 is highly expressed following the degeneration of SGNs. Finally, we observed that the increased SDF-1 is derived from the Schwann cells in the SGN-degenerated model. These results suggest that SDF-1, which is derived from cochlear Schwann cells and up-regulated in the early injury microenvironment, plays a beneficial role in the NSC migration to the injury region. Optimizing SDF-1 expression in the host microenvironment or increasing the CXCR4 expression of the donor stem cells may improve the migration efficiency of transplanted cells toward the injury region in the cochlea., (Copyright © 2012 Elsevier Ireland Ltd. All rights reserved.)

Objective: Recurrent aphthous stomatitis (RAS) is a common oral mucosal disease, yet effective therapeutic approaches are lacking. This study aimed to determine the effects of application of berberine gelatin in the treatment of minor RAS (MiRAS)., Methods: A randomized, double-blind, placebo-controlled, clinical trial was performed. The gelatin containing berberine (5 mg/g) or vehicle only was applied 4 times per day for 5 days. Clinical evaluation included pain level, size, erythema, and exudation of certain ulcers on days 1, 2, 4, and 6., Results: A total of 84 subjects fulfilled the study without obvious side effects. Berberine gelatin treatment reduced the ulcer pain score compared with placebo gelatin (P < 0.05). Ulcer size was significantly reduced (P < 0.05) and lower erythema (P < 0.05) and exudation (P < 0.05) levels were associated with berberine treatment., Conclusions: Berberine gelatin may be a safe and effective treatment for MiRAS., (Copyright © 2013 Elsevier Inc. All rights reserved.)

Diethylstilbestrol (DES) is a nonsteroidal synthetic estrogen widely used in estrogen therapy. In animal models, exposure to DES disrupts the outgrowth of the gubernacula, leading to testis maldescent. However, it remains unclear whether the effects of DES on gubernaculum are direct or indirect, and the underlying mechanisms are largely obscure. In this study, mouse gubernaculum testis cells were isolated and treated by DES, and cell morphology and function were examined. The results showed that DES changed the morphology and inhibited the proliferation of gubernacular cells. Furthermore, DES increased intracellular [Ca(2+)] and induced F-actin rearrangement and stress fiber formation in gubernaculum testis cells in a dose-dependent manner. Taken together, these data suggest that DES impairs the morphology and inhibits the proliferation and contractility of gubernaculum testis cells. The experimental model we established and our observations based on this model help provide new insight into the role of DES in the etiology of cryptorchidism.

Objective: To investigate the impact of prenatal exposure to diethylstilbestrol (DES) on the specific receptor LGR8 of insulin-like factor 3 (INSL3) in the mouse gubernaculum testis, and that of exoestrogens on descensus testis in mice., Methods: A total of 120 pregnant KM mice aged 8 to 10 weeks were assigned to a normal, a blank control and 4 DES groups of equal number, the blank controls injected subcutaneously with dimethyl sulfoxide plus normal saline, and the DES groups with DES at 0.1, 1, 10 and 100 microg/kg body weight, respectively, from embryonic day 9 (ED9) through ED17. Immunohistochemistry and RT-PCR were used to detect the expressions of LGR8 protein and mRNA in the gubernaculum testis of the ED18 fetuses and PND20 (postnatal day 20) offspring of the mice., Results: Histological analysis showed that the gubernaculum testis of the ED18 fetuses were well developed in both the normal and control groups, with an inner mesenchymal core and muscular outer layer. In contrast, the gubernaculum testis were poorly developed in the experimental groups, morphologically abnormal and without visible dividing line between the mesenchymal tissue and the muscular outer layer. No obvious differences were found in the gubernaculum testis development of the neonates between the normal and experimental groups. Positive immunostaining was seen in the mesenchymal core and muscular outer layer, but mainly in the latter. The expression of LGR8 was weaker in the experimental groups than in the normal group (P < 0.05), but that of LGR8 mRNA was increased in the high-dose (10 and 100 microg/kg) DES groups (P < 0.05). No obvious mutations were observed in the PCR products in any of the experimental groups., Conclusion: Prenatal exposure to diethylstilbestrol affected the expression of LGR8 mRNA in the mouse gubernaculum testis, which suggests that diethylstilbestrol may induce cryptorchidism by interfering with the INSL3-LGR8 signaling system and consequently the development of the gubernaculum testis.

Objective: We investigated the effectiveness and safety of topical application of 5 mg allicin adhesive tablets in the treatment of minor recurrent aphthous ulcerations (MiRAU)., Material and Methods: A randomized, double-blinded, placebo-controlled, clinical trial was performed. Tablets containing 5 mg allicin or vehicle only were consecutively applied 4 times per day for 5 days. The size and pain level of ulcers were measured and recorded on days 1, 2, 4, and 6., Results: A total of 96 subjects with MiRAU fulfilled the study. Allicin adhesive tablets significantly reduced ulcer size (P < .005, P < .003, P < .001 for days 2, 4, and 6, respectively) and alleviated ulcer pain score (P < .03, P < .001, P < .05 for days 2, 4, and 6, respectively) compared with vehicle tablets. Minor and major adverse reactions were not observed., Conclusions: Allicin adhesive tablets were effective in reducing ulcer size and alleviating ulcer pain of the patients in the treatment of MiRAU without significant side effects., (Crown Copyright © 2012. Published by Mosby, Inc. All rights reserved.)

Pelagibacterium halotolerans B2(T) is a marine halotolerant bacterium that was isolated from a seawater sample collected from the East China Sea. Here, we present the complete genome sequence of the type strain P. halotolerans B2(T), which consists of one chromosome (3,944,837 bp; 61.4% G+C content) and one plasmid (4,050 bp; 56.1% G+C content). This is the first complete genome of a member of the Pelagibacterium genus.

Cobalt-rich crusts are important metallic mineral resources with great economic potential, usually distributed on seamounts located in the Pacific Ocean. Microorganisms are believed to play a role in the formation of crusts as well as in metal cycling. To explore the microbial diversity related to cobalt-rich crusts, 16S ribosomal RNA gene clone libraries were constructed from three consecutive sediment layers. In total, 417 bacterial clones were obtained from three bacterial clone libraries, representing 17 distinct phylogenetic groups. Proteobacteria dominated in the bacterial communities, followed by Acidobacteria and Planctomycetes. Compared with high bacterial diversity, archaea showed a remarkably low diversity, with all 137 clones belonging to marine archaeal group I except one novel euryarchaeotal clone. The microbial communities were potentially involved in sulfur, nitrogen and metal cycling in the area of cobalt-rich crusts. Sulfur oxidation and metal oxidation were potentially major sources of energy for this ecosystem. This is the first reported investigation of microbial diversity in sediments associated with cobalt-rich crusts, and it casts fresh light on the microbial ecology of these important ecosystems., (© 2011 Federation of European Microbiological Societies. Published by Blackwell Publishing Ltd. All rights reserved.)

A new agarase, AgaA(CN41), cloned from Vibrio sp. strain CN41, consists of 990 amino acids, with only 49% amino acid sequence identity with known β-agarases. AgaA(CN41) belongs to the GH50 (glycoside hydrolase 50) family but yields neoagarotetraose as the end product. AgaA(CN41) was expressed and characterized.

To cost-efficiently produce biofuels, new methods are needed to convert lignocellulosic biomass into fermentable sugars. One promising approach is to degrade biomass using cellulosomes, which are surface-displayed multicellulase-containing complexes present in cellulolytic Clostridium and Ruminococcus species. In this study we created cellulolytic strains of Bacillus subtilis that display one or more cellulase enzymes. Proteins containing the appropriate cell wall sorting signal are covalently anchored to the peptidoglycan by coexpressing them with the Bacillus anthracis sortase A (SrtA) transpeptidase. This approach was used to covalently attach the Cel8A endoglucanase from Clostridium thermocellum to the cell wall. In addition, a Cel8A-dockerin fusion protein was anchored on the surface of B. subtilis via noncovalent interactions with a cell wall-attached cohesin module. We also demonstrate that it is possible to assemble multienzyme complexes on the cell surface. A three-enzyme-containing minicellulosome was displayed on the cell surface; it consisted of a cell wall-attached scaffoldin protein noncovalently bound to three cellulase-dockerin fusion proteins that were produced in Escherichia coli. B. subtilis has a robust genetic system and is currently used in a wide range of industrial processes. Thus, grafting larger, more elaborate minicellulosomes onto the surface of B. subtilis may yield cellulolytic bacteria with increased potency that can be used to degrade biomass.

46, XY disorders of sexual development (46, XY DSD) are a group of complicated clinical conditions, which involve medical care, society, ethics and many other aspects. As chronic diseases, they necessitate long-term or even lifelong physical and mental follow-up and treatment. Early diagnosis and reasonable treatment could not only achieve appropriate development of the secondary sexual characteristics, but also effectively prevent gonadal malignancy. In recent years, environment pollution and other factors are contributing to the increasing incidence of 46, XY DSD all over the world. A deeper clinical insight into these disorders helps their earlier diagnosis and maximum improvement of prognosis. The etiology, new classification and treatment of 46, XY DSD are reviewed in this article.

In the present study, rat model of gastric ischemia-reperfusion (GI-R) injury was established by clamping the celiac artery for 30 min followed by 1 h of reperfusion. Subsequently, the regulatory effect of electrical stimulation of cerebellar fastigial nucleus (FN) on GI-R injury and its neural mechanisms were investigated in Sprague-Dawley rats. The results are as follows. Electrical stimulation of the cerebellar FN not only obviously attenuated the GI-R injury in an intensity-dependent manner, but also decreased the apoptosis rate of gastric mucosal cells. Chemical lesion of FN eliminated the protective effect of electrical stimulation of FN on GI-R injury. Electrical stimulation of cerebellar FN decreased both the frequency and amplitude of the discharges of greater splanchnic nerve, but it could not change the discharge of greater splanchnic nerve following the lesion of the lateral hypothalamic area (LHA). After bilateral section of the greater splanchnic nerves, electrical stimulation of the FN also attenuated the GI-R injury. Chemical lesion of the LHA reversed the protective effect of electrical stimulation of FN on GI-R injury. Electrical stimulation of FN increased the activity of superoxide dismutase (SOD), but decreased the content of malondialdehyde (MDA) in gastric mucosa under GI-R. These results indicate that the cerebellar FN may regulate GI-R injury. Therefore, the cerebellar FN is an important brain site protecting the stomach against GI-R. The LHA and greater splanchnic nerves participate in the regulatory effects of cerebellar FN stimulation on GI-R injury. In addition, antioxidation may also be involved in the protection mechanism of cerebellar FN stimulation.

Objective: To establish a primary culture of the testis gubernacular cells of Kunming mice, observe the morphological characteristics of the cells, and explore the effects of exogenous estrogens (EEs) on the development of the testis gubernacula in vitro., Methods: We removed the gubernacula from 3-day-old mice with the surgical magnifier and cultured the gubernacular cells. Then we detected the cell viability by trypan blue and cell morphology by HE staining. The subcultured cells were randomly divided into a blank control, a DMSO (0.1%, v/v) control, and 4 experimental groups (given 0.01, 0.10, 1.00 and 10.00 micdrog/ml of diethylstilbestrol [DES] dissolved in DMSO, respectively). After treated for 12, 24 and 48 hours, the gubernacular cells were observed for morphological changes and proliferation inhibition by CCK-8., Results: Most of the cultured gubernacular cells were fibroblasts, and a few were epithelioids. The primary cells showed a viability of 85%-90%. Dose- and time-dependent inhibition of cell proliferation was found in the four experimental groups at three different times, with statistically significant differences (P < 0.01)., Conclusion: Gubernacular cells can be cultured in vitro. EEs inhibit the proliferation of gubernacular cells in a dose- and time-dependent manner. An in- sight into the effects EES on cultured gubernacular cells is an effective approach to the study of their influence on the development of the reproductive system.

Pandoraea sp. strain MCB032 was isolated as an emerging chlorobenzene degrader from a functionally stable bioreactor where species succession had occurred. In this study, two gene clusters encoding chlorobenzene metabolic functions have been cloned. Within the cbs gene cluster, CbsA and CbsB are similar to the chlorobenzene dioxygenase and the cis-chlorobenzene dihydrodiol dehydrogenase in Ralstonia sp. JS705 and shown to transform chlorobenzene to 3-chlorocatechol. The clc gene cluster shows strong similarity to the clc genes of Ralstonia sp. JS705 and encodes chlorocatechol 1,2-dioxygenase (ClcA) and other enzymes, which catalyze the conversion of chlorocatechol to 3-oxoadipate. The Michaelis constants (K (m)) values of ClcA for catechol, 3-methylcatechol and 3-chlorocatechol were determined as 10.0, 8.9 and 3.4 muM, respectively. CbsX, a putative transport protein present in the cbs cluster of strain MCB032 but not in those of other chlorobenzene degraders, shows 76 and 53% identities to two previously identified transport proteins involved in toluene degradation, TbuX from Ralstonia pickettii PKO1 and TodX from Pseudomonas putida F1. The presence of the transport protein in strain MCB032 likely provides a mechanistic explanation for its higher chlorobenzene affinity and may well be the basis for the competitive advantage of this strain in the bioreactor.

Objective: To investigate the role of nasal mucosa fibrosis on radiation induced nasal mucosa injury., Methods: Seventy two male rats were randomly divided into two groups, control group and irradiation injured group (radiation dose were 40 Gy); the rats were killed 1 week, 2 weeks, 4 weeks, 8 weeks, 3 months and 6 months after the finish of radiation. The middle turbinates of the animals were removed. The pathological change of the nasal mucosa were observed with scanning electron microscope, transmission electron microscope, hematoxylin and eosin (HE), alcian blue-periodic acid-Schif (AB-PAS), and Masson Trichrome (MT). The Hyp content in nasal mucosa was measured with chemo-chromatometry., Results: After radiation, the pathological characteristics in early stage (within 4 weeks) was acute inflammatory reaction. The repair of nasal mucosa started 4 weeks after radiation, lasted to 6 months. The deposition of collagen in nasal mucosa could be found 1 week after irradiation and increased gradually., Conclusion: Irradiation could induce a serials of pathological changes on nasal mucosa. The nasal mucosa fibrosis may be one of the reasons of persistent irradiation induced nasal mucosa injury.

Recurrent aphthous ulcer (RAU) is the most common ulcer of oral non-keratinized mucosa, but the treatment is always limited at present. Considering the multifactorial etiology of RAU, a novel therapeutic agent with multi-bioactivties should be presented. Garlic has been recognized as a favorable natural medicine against a large number of pathologic conditions. The major component of garlic is allicin, which could effectively decrease inflammatory factors secretion, reduce the migration of neutrophils, inhibit bacterium and virus, antagonize oxidation and regulate immunity. By these bioactivities of anti-inflammation, anti-microbial activity, anti-oxidation and immunomodulation, the allicin may be an effective therapeutic candidate to control the pain, promote ulcer healing and prevent the recurrence of RAU.

Objective: To explore the feasibility of serial slices microscopic histological investigation for the elaborate evaluation of reproductive system malformations., Methods: Newborn male mice prenatally exposed to different doses of subcutaneously given diethylstilbestrol (DES) from gestational day 9 to 17 were treated by fixing parts of the abdomen in situ and setting them to transected serial slices. All the slices were stained, studied under the microscope and serially recorded by software. The gubernaculum was morphologically analyzed and its location and size were measured., Results: Morphologically, the gubernaculum could be identified clearly, its structure inhomogeneous from proximal to distal and dissymmetric from right to left. The environmental estrogen produced different effects on the morphology of the gubernaculum in different parts and most obviously affected its length., Conclusion: Prenatal exposure to environmental estrogen has evident and general effects on the gubernacular development of newborn male mice. The morphological study with serial histological slices gives a precise and systematic evaluation of genital malformations.

Aim: To investigate the role of overexpression of beta2-adrenoceptor on contraction in cardiac myocytes isolated from failure hearts of rats and primarily analyses its mechanisms., Methods: Primarily cultured cardiac myocytes were infected with adenovirus containing the sequence for human beta2-adrenoceptors. The expression of beta2-adrenoceptors was tested by Western blot. The contraction amplitudes induced by isoprenaline stimulation were measured., Results: Overexpression of beta2-adrenoceptor increased the content in failure cardiac myocytes. The contraction amplitudes in failure cardiac myocytes were lower than that in the control (P < 0.01). Overexpression of beta2 adrenoceptor improved the contraction of failure cardiac myocytes (P < 0.01, Failure+ Adv.Beta2 group vs. Failure group). Selective beta2-adrenoceptor antagonist ICI 118,551 partially reversed the effects (P < 0.05, Failure+ Adv.beta2 + ICI group vs Failure + Adv.beta2 group), but the contraction amplitudes in this Failure +/- Adv.beta2 + ICI 118,551 group were still higher than that in only heart failure group (P < 0.05). Selective beta1 adrenoceptor antagonist CGP20712A completely inhibited the effects of overexpression of beta2 adrenoceptor on contraction amplitude in failure cardiac myocytes., Conclusion: Overexpression of beta2-adrenoceptors improves the contraction of cardiac myocytes isolated from failure hearts of rats. The effect is related to beta1-adrenoceptor.

Objective: To investigate the isolation and expansion of mesenchymal stem cells (MSCs) from human umbilical cord Wharton's jelly and their biological identities, and explore the possibility of inducing human umbilical cord-derived MSCs to differentiate into neurocyte-like cells., Methods: The growth and proliferative abilities of human umbilical cord-derived MSCs were observed, and their immunophenotypes were determined by flow cytometry. Salvia miltiorrhiza and beta-sulfhydryl alcohol were adopted to induce the cells to differentiate. The differentiated and undifferentiated cells were identified with immunocytochemistry. The pleiotrophin and nestin genes were measured by RT-PCR., Results: A population of human umbilical cord-derived MSCs were isolated from human umbilical Wharton's jelly; they were processed to obtain a fibroblast-like population of cells and could be maintained in vitro for extended periods with stable population doubling, and they were expanded as undifferentiated cells in culture for more than 10 passages, indicating their proliferative capacity. The human umbilical cord-derived MSCs were positive for CD(29), CD(44), CD(59), CD(105), but negative or weakly expressed the markers of hematopoietic cells such as CD(14), CD(33), CD(34), CD(28), CD(45) and CD(117). The important GVHD correlation markers were negative or weakly expressed, including CD(80) (B7-1), CD(86) (B7-2), CD(40) and CD(40L). Salvia miltiorrhiza beta-sulfhydryl alcohol could induce the MSCs to express nestin, a marker of neuronal precursor stem cells at early stage of differentiation. Later, they exhibited neural phenotypes, expressing beta-tubulin III and neurofilament (NF) and glial fibrillary acidic protein (GFAP). It was confirmed by RT-PCR that the MSCs could express pleiotrophin either before or after the induction of salvia miltiorrhiza, furthermore, after the induction the expression was markedly enhanced and the nestin gene was also expressed., Conclusion: The human MSCs could be isolated from human umbilical cord Wharton's jelly, and it was easy to propagate these MSCs. The negative GVHD correlated markers might result from the fact that MSCs had no HLA barrier, which may suggest potential clinical significance. The MSCs are capable of differentiating into neurocyte-like cells and they may represent an alternative stem cell source for CNS cells transplantation.

The testicular gubernaculum plays an important role in testicular descent and development. Its differentiation and development are affected by many factors. Androgens, calcitonin gene-related peptide (CGRP), insulin-like factor 3 (INSL3), Müllerian inhibiting substance (MIS), epidermal growth factor (EGF) and environmental estrogens (EEs) are involved in gubernacular development. The effect of CGRP, INSL3 and especially EEs on genital system has been attracted more attention.

Testicular descent is an essential step in the course of reproductive system development. The mechanisms involved in the regulation of testis descent is not distinct. Gubernaculum has a very close relationship with testis descent. Maldescent of testis can cause abnormalities of genital system such as testicular underwent (cryptorchidism), dysplasia, tumor, infertility and low sexuality. Recently insulin like hormone 3 is a hotspot of concerning affecting gubernacular development and testicular descent. This article briefly reviews the advances in these aspects.

Background: The two most basic properties of mesenchymal stem cells (MSCs) are the capacities to self-renew indefinitely and differentiate into multiple cells and tissue types. The cells from human umbilical cord Wharton's Jelly have properties of MSCs and represent a rich source of primitive cells. This study was conducted to explore the possibility of inducing human umbilical cord Wharton's Jelly-derived MSCs to differentiate into nerve-like cells., Methods: MSCs were cultured from the Wharton's Jelly taken from human umbilical cord of babies delivered after full-term normal labor. Salvia miltiorrhiza and beta-mercaptoethanol were used to induce the human umbilical cord-derived MSCs to differentiate. The expression of neural protein markers was shown by immunocytochemistry. The induction process was monitored by phase contrast microscopy, electron microscopy (EM), and laser scanning confocal microscopy (LSCM). The pleiotrophin and nestin genes were measured by reverse transcription-polymerase chain reaction (RT-PCR)., Results: MSCs in the Wharton's Jelly were easily attainable and could be maintained and expanded in culture. They were positive for markers of MSCs, but negative for markers of hematopoietic cells and graft-versus-host disease (GVHD)-related cells. Treatment with Salvia miltiorrhiza caused Wharton's Jelly cells to undergo profound morphological changes. The induced MSCs developed rounded cell bodies with multiple neurite-like extensions. Eventually they developed processes that formed networks reminiscent of primary cultures of neurons. Salvia miltiorrhiza and beta-mercaptoethanol also induced MSCs to express nestin, beta-tubulinIII, neurofilament (NF) and glial fibrillary acidic protein (GFAP). It was confirmed by RT-PCR that MSCs could express pleiotrophin both before and after induction by Salvia miltiorrhiza. The expression was markedly enhanced after induction and the nestin gene was also expressed., Conclusions: MSCs could be isolated from human umbilical cord Wharton's Jelly. They were capable of differentiating into nerve-like cells using Salvia miltiorrhiza or beta-mercaptoethanol. The induced MSCs not only underwent morphologic changes, but also expressed the neuron-related genes and neuronal cell markers. They may represent an alternative source of stem cells for central nervous system cell transplantation.

Background: Hemangiomas are the most common tumors in children. Some hemangiomas may require intervention because of their location, size, behavior, or potential for important complications. Pharmacological therapy with glucocorticoids is the mainstay treatment, but there is no consensus on therapeutic regimens or candidate selection, therapeutic efficacy varies, and the mechanism mediating the beneficial effects of glucocorticoids remains unclear. This study was performed to investigate the expression patterns of the glucocorticoid receptor (GR) and its alpha isoform (GRalpha) in cutaneous hemangiomas and vascular malformations., Methods: SP immunohistochemical technique was used to examine the expression of GR(e-20) (GR) and GR(p-20) (GRalpha) on vascular endothelial cells in 80 specimens that included 33 proliferating hemangiomas, 32 involuting hemangiomas, 7 vascular malformations as well as 8 normal skin tissues, all obtained from infants and children. GR and GRalpha expression in prepared tissue slides were examined using automated computer-assisted microscopic analysis. Mean gray scale values were compared among the various tumor types., Results: The mean gray scale values of GR were 127.0 +/- 6.4 and 121.4 +/- 6.6 in hemangiomas and vascular malformations respectively, but this difference was not statistically significant (P = 0.104). However, these values were all markedly higher than that of normal skin, which was only 108.6 +/- 6.8 (P = 0.001 and P = 0.000 for comparison with hemangiomas and vascular malformations respectively). The gray scale of GR in proliferation and involuting hemangiomas were 127.9 +/- 4.8 and 126.0 +/- 5.8 respectively, but this difference was not significant (P = 0.146). However, GRalpha expression in hemangiomas, vascular malformations and normal skin declined gradually in stepwise fashion (127.3 +/- 5.4, 120.4 +/- 6.1 and 109.9 +/- 5.3 respectively; P < 0.001). GRalpha expression was higher in proliferating hemangiomas than in involuting hemangiomas (127.2 +/- 6.3 and 122.5 +/- 6.3; P = 0.004)., Conclusions: GR and GRalpha are strongly expressed in hemangiomas and vascular malformations. The expression of GRalpha is closely related to the phase of the hemangioma. Determination of GR and GRalpha may be a positive significance to understand the information of hemangiomas and vascular malformations and may further help determining proper strategies of steroid therapy for hemangiomas and vascular malformations.

We report on an optical interference method for transferring periodic microstructures of metal film from a supporting substrate to a receiving substrate by means of five-beam interference of femtosecond laser pulses. Scanning electron microscopy and optical microscopy revealed microstructures with micrometer-order were transferred to the receiving substrate. In the meanwhile, a negative copy of the transferred structures was induced in the metal film on the supporting substrate. The diffraction characteristics of the transferred structures were also evaluated. The present technique allows one-step realization of functional optoelectronic devices.

Writing computer-generated holograms have been achieved by using a near infrared femtosecond laser selective ablation of metal film deposited on glass substrate. The diffraction features with data reconstruction of the fabricated computer-generated holograms were evaluated. Both transmission and reflection holograms can be fabricated in a single process. The process requires no mask, no pre- or post-treatment of the substrate.

We report near infrared broadband emission of bismuth-doped barium-aluminum-borate glasses. The broadband emission covers 1.3microm window in optical telecommunication systems. And it possesses wide full width at half maximum (FWHM) of ~200nm and long lifetime as long as 350micros. The luminescent properties are quite sensitive to glass compositions and excitation wavelengths. Based on energy matching conditions, we suggest that the infrared emission may be ascribed to 3P1? 3P0 transition of Bi+. The broad infrared emission characteristics of this material indicate that it might be a promising candidate for broadband optical fiber amplifiers and tunable lasers.

Near infrared broadband emission characteristics of bismuth-doped aluminophosphate glass have been investigated. Broad infrared emissions peaking at 1210nm, 1173nm and 1300nm were observed when the glass was pumped by 405nm laser diode (LD), 514nm Ar+ laser and 808nm LD, respectively. The full widths at half maximum (FWHMs) are 235nm, 207nm and 300nm for the emissions at 1210nm, 1173nm and 1300nm, respectively. Based on the energy matching conditions, it is suggested that the infrared emission may be ascribed to 3P1? 3P0 transition of Bi+. The broadband infrared luminescent characteristics of the glasses indicate that they are promising for broadband optical fiber amplifiers and tunable lasers.

Aim: To study the effect of prenatal exposure to diethylstilbestrol (DES) and the role of actin and proliferating cell nuclear antigen (PCNA) on testicular gubernaculum development in fetal male Kunming mice., Methods: Pregnant mice were randomly assigned to 6 groups and injected with DES subcutaneously from gestational day 9 (E9) to day 17 (E17) at doses of 0, 25, 50, 100, 200 microg.kg-1.d-1 in 0.2 mL dimethyl sulfoxide (DMSO). On E17 they were sacrificed and fetuses quickly removed for fixation. Male fetuses were sliced on serial coronal plane. Histological changes were observed under the light microscope (LM) and ultrastructural changes with the scanning and transmission electron microscopes (SEM and TEM). The expression intensity of actin and PCNA in the gubernacula was quantitated by immunohistochemistry., Results: The mortality of the fetuses was higher in the DES-treated groups than that in the DMSO and saline controls (P<0.05). Under LM the gubernacula were seen to be poorly developed with smaller bulbs. On SEM the bulbs lose the clear demarcation between the mesenchymal inner core and the muscular outer layer and looked like a small cone instead of the normal cylindrical appearance. On TEM there were some smaller disordered myofibrils and sparse cytoplasmic organelles in the gubernacular muscular cells of the treated groups. The expression intensity of actin and PCNA in the gubernacula was significantly weaker in the treated groups than that in the DMSO and saline controls (P<0.05)., Conclusion: DES induces underdevelopment of the gubernacula in a dose-dependent manner in fetal male mice and down regulates the actin and PCNA expression.

We report a controllable process of recipitation and dissolution of silver nanoparticles in ultrashort laser pulses irradiated Ag+-doped phosphate glass. Absorption spectra, transmission electron microscopy and refractive index measurement revealed that metallic silver nanoparticles were precipitated in the glass sample after irradiation by an 800-nm femtosecond laser and subsequent annealing at 300 degrees C, and dissolved after further annealing at 450 degrees C. We discuss a mechanism that combines the formation and decoloration of color centers, precipitation and dissolution of silver nanoparticles.

We report the fabrication of internal diffraction gratings in calcium fluoride crystals by a focused near-IR 800 nm femtosecond laser. The diffraction efficiency and refractive index change were evaluated after femtosecond laser irradiation and subsequent annealing. The maximum refractive index change was estimated to be 3.57x10(-4). Optical absorption spectra, measured for the crystals before and after the laser irradiation and subsequent annealing, indicate that the absorbance increase after femtosecond laser irradiation and decrease with increasing annealing temperature. The mechanisms of refractive index change are proposed. The results may be useful for fabrication of three-dimensional integrated optics devices in the crystals.

Objective: To search for a sedative method that is more suitable for gastroscopy., Methods: All of patients were randomly divided into control group and experimental group. The experimental group was treated with propofol and midazolam, the control group was treated with propofol alone. The cumulation dosage of propofol, sedative effect, variation of BP and SaO2 were observed in all patient., Results: The cumulative dosage of propofol in the experimental group was lower than that in the control group [(73.21 +/- 18.67) mg and (117.23 +/- 21.57) mg respectively]; the oblivious degree in the experimental group was higher than that in the control group (95.65% and 80.00%); the onset time and the descendant range of BP and SaO2 were also lower in the experimental than those in the control group. There was not remarkable difference in sedative effect and veriviscont time between the control group and the experimental group., Conclusions: In such a rapid operation of gastroscopy, the dosage of propofol in the experimental group is obviously less than that in the control group, while it does not affect the effect of sedation, the diagnose and cure time in gastroscopy room, and has more security and less cost.

We test the hypothesis that microvascular endothelial cells may undergo apoptosis in response to acute pulmonary venous hypertension. The isolated rabbit lungs were perfused in situ for 4 h with left atrial pressure of 0, 10, or 20 mmHg at a constant blood flow. Edema formation was monitored by lung weight gain. To assay for apoptosis, we performed agarose gel electrophoresis of DNA, in situ nick end labeling of DNA strand breaks, and electron microscopy. We also examined the levels of expression of Bcl-2, a suppressor of apoptosis, in microvascular endothelial cells using an immunohistochemical technique. In a vascular pressure-dependent fashion, we found apoptosis in endothelial cells of alveolar septal capillaries, as well as expression of Bcl-2 in arteriolar and venular endothelial cells. We conclude that acute pulmonary venous hypertension induces apoptosis in capillary endothelial cells but not in arteriolar and venular endothelial cells, suggesting that microvascular endothelial cell apoptosis is dependent on the levels of Bcl-2 expression and influences the formation or resolution of acute hydrostatic lung edema.

Background: Helicobacter pylori, especially the CagA-positive strains, are closely associated with peptic ulcers and gastric cancers. We performed a large scale gastric cancer screening project and examined the prevalence of H. pylori and CagA-positive strains in Changle, China, an area with one of the World's highest gastric cancer mortality. We also compared the prevalence with that in Hong Kong which has one-tenth of the gastric cancer mortality of that in Changle., Methods: A total of 2424 subjects in Changle and 523 subjects in Hong Kong had endoscopic examination and venesection. Sera were tested for anti-H. pylori antibody and anti-CagA antibody and correlated with endoscopic findings., Results: In Changle, 80. 9% of the subjects were H. pylori carriers. Out of 551 carriers, 408 (74%) were positive for anti-CagA antibody. A total of 76% and 87% of the asymptomatic and gastric cancer patients were positive for anti-CagA antibody, respectively (P > 0.05). Compared to Hong Kong, there was a significantly (P < 0.0001) higher prevalence of CagA-positive strains in asymptomatic subjects in Changle (76%) than in Hong Kong (28%), but not in peptic ulcers or gastric cancers., Conclusions: Subjects in Changle had a high prevalence of H. pylori infection and a high prevalence of the CagA-positive strains. The contrast in the prevalence of CagA-positive strains, in asymptomatic subjects in two areas with differing gastric cancer mortality, supports the pathogenic role of CagA-positive strains in gastric carcinogenesis.

Background: Carriers of Helicobacter pylori are believed to have a three- to six-fold increased risk of developing gastric cancer. We have recently conducted a simultaneous cross-sectional population study on the prevalence of H. pylori infection in a cohort of asymptomatic adult volunteers in two contrasting gastric cancer risk regions of South China, Hong Kong and Changle of Fujian. Their mean annual gastric cancer mortality has been approximately 7.5 and 75/100 000 population, respectively, since the beginning of the last decade. The aim of this study was to evaluate if H. pylori prevalence bears any relationship to gastric cancer mortality rates in these two southern regions of China., Methods: Sera were obtained from 397 volunteers in Hong Kong. They were tested for anti-H.pylori immunoglobulin (Ig) G antibody by using a commercial enzyme-linked immunosorbent assay (ELISA) kit. Volunteers of Changle (1456) had upper endoscopy examination and were also tested for anti-H. pylori IgG antibody by the same ELISA method., Results: The overall H. pylori infection prevalence was significantly higher in Changle (80.4%) than in Hong Kong (58.4%; P< 0.01). The high prevalence is associated with more atrophic gastritis. The overall risk of gastric cancer in people of Changle is approximately five-fold that of Hong Kong (adjusted odds ratio 4.9, 95% CI 2.5-9.8)., Conclusions: It is concluded that the prevalence of H. pylori infection rates bear a direct relationship to gastric cancer mortality rates in these two southern regions of China. Thus, H. pylori most likely plays a significant aetiopathogenetic role in gastric carcinogenesis in subjects living in Changle.