Exosomes combined with biosynthesized cellulose conduits improve peripheral nerve regeneration.

Peripheral nerve injury is one of the more common forms of peripheral nerve disorders, and the most severe type of peripheral nerve injury is a defect with a gap. Biosynthetic cellulose membrane (BCM) is a commonly used material for repair and ligation of nerve defects with gaps. Meanwhile, exosomes from mesenchymal stem cells can promote cell growth and proliferation. We envision combining exosomes with BCMs to leverage the advantages of both to promote repair of peripheral nerve injury. Prepared exosomes were added to BCMs to form exosome-loaded BCMs (EXO-BCM) that were used for nerve repair in a rat model of sciatic nerve defects with gaps. We evaluated the repair activity using a pawprint experiment, measurement and statistical analyses of sciatica function index and thermal latency of paw withdrawal, and quantitation of the number and diameter of regenerated nerve fibers. Results indicated that EXO-BCM produced comprehensive and durable repair of peripheral nerve defects that were similar to those for autologous nerve transplantation, the gold standard for nerve defect repair. EXO-BCM is not predicted to cause donor site morbidity to the patient, in contrast to autologous nerve transplantation. Together these results indicate that an approach using EXO-BCM represents a promising alternative to autologous nerve transplantation, and could have broad applications for repair of nerve defects.

[Low-intensity pulsed ultrasound versus low-energy shock wave in the treatment of neurogenic penile erectile dysfunction].

OBJECTIVE: To investigate the effect and safety of low-intensity pulsed ultrasound (LIPUS) versus low-energy shock wave (LESW) in the treatment of neurogenic penile ED in male SD rats. METHODS: Twenty-four 12-week-old male SD rats were randomly divided into four groups of an equal number: sham operation, bilateral cavernous nerves injury (BCNI), LIPUS (300 mW /cm2, 3 times a week for 2 weeks), and LESW (300 strokes once, 3 times a week for 2 weeks). At 28 days after intervention, the erectile function of the rats was assessed by comparing the ratio of maximum intracavernous pressure to mean arterial pressure (ICPmax/MAP), and the histopathological changes in the corpus cavernosum of the penis were observed by immunohistochemistry, immunofluorescence staining, and Masson trichromatic staining. RESULTS: After treatment, the LIPUS and LESW groups, compared with the BCNI group, showed significantly increased ICPmax/MAP ratio (0.56 ± 0.13 and 0.55 ± 0.10 versus 0.35 ± 0.14, P = 0.017 and P = 0.013), improved smooth muscle/collagen value (0.08 ± 0.01 and 0.08 ± 0.02 versus 0.06 ± 0.02, P = 0.017 and P = 0.019), and elevated proportion of smooth muscle to cavernosum (0.20 ± 0.05 and 0.21 ± 0.03 versus 0.15 ± 0.02, P = 0.046 and P = 0.020), with no statistically significant difference between the LIPUS and LESW groups. No obvious adverse reactions were observed in the LIPUS or LESW group. CONCLUSIONS: Both LIPUS and LESW can effectively improve penile erectile function and repair histopathological injury in the animal model of neurogenic ED.

Involvement of abscisic acid-responsive element-binding factors in cassava (Manihot esculenta) dehydration stress response.

Cassava (Manihot esculenta) is a major staple food, animal feed and energy crop in the tropics and subtropics. It is one of the most drought-tolerant crops, however, the mechanisms of cassava drought tolerance remain unclear. Abscisic acid (ABA)-responsive element (ABRE)-binding factors (ABFs) are transcription factors that regulate expression of target genes involved in plant tolerance to drought, high salinity, and osmotic stress by binding ABRE cis-elements in the promoter regions of these genes. However, there is little information about ABF genes in cassava. A comprehensive analysis of Manihot esculenta ABFs (MeABFs) described the phylogeny, genome location, cis-acting elements, expression profiles, and regulatory relationship between these factors and Manihot esculenta betaine aldehyde dehydrogenase genes (MeBADHs). Here we conducted genome-wide searches and subsequent molecular cloning to identify seven MeABFs that are distributed unevenly across six chromosomes in cassava. These MeABFs can be clustered into three groups according to their phylogenetic relationships to their Arabidopsis (Arabidopsis thaliana) counterparts. Analysis of the 5'-upstream region of MeABFs revealed putative cis-acting elements related to hormone signaling, stress, light, and circadian clock. MeABF expression profiles displayed clear differences among leaf, stem, root, and tuberous root tissues under non-stress and drought, osmotic, or salt stress conditions. Drought stress in cassava leaves and roots, osmotic stress in tuberous roots, and salt stress in stems induced expression of the highest number of MeABFs showing significantly elevated expression. The glycine betaine (GB) content of cassava leaves also was elevated after drought, osmotic, or salt stress treatments. BADH1 is involved in GB synthesis. We show that MeBADH1 promoter sequences contained ABREs and that MeBADH1 expression correlated with MeABF expression profiles in cassava leaves after the three stress treatments. Taken together, these results suggest that in response to various dehydration stresses, MeABFs in cassava may activate transcriptional expression of MeBADH1 by binding the MeBADH1 promoter that in turn promotes GB biosynthesis and accumulation via an increase in MeBADH1 gene expression levels and MeBADH1 enzymatic activity. These responses protect cells against dehydration stresses by preserving an osmotic balance that enhances cassava tolerance to dehydration stresses.

The American cockroach peptide periplanetasin-4 inhibits Clostridium difficile toxin A-induced cell toxicities and inflammatory responses in the mouse gut.

Many reports have shown that crude extracts of the American cockroach have therapeutic effects on inflammation. In a previous study, our research group showed that an antimicrobial peptide (Periplanetasin-2) derived from the American cockroach via de novo transcriptome analysis inhibited apoptosis of human colonocytes and inflammatory responses of the mouse gut caused by Clostridium difficile toxin A. Here, we examined whether Periplanetasin-4 (Peri-4), another antimicrobial peptide identified via de novo transcriptome analysis of the American cockroach, could also inhibit the various toxicities induced by C. difficile toxin A. We found that Peri-4 significantly reduced the cell viability loss and cell apoptosis caused by toxin A in vitro. Peri-4 also ameliorated the severe inflammatory responses seen in the toxin A-induced mouse enteritis model, rescuing the villus disruption and interleukin-6 production induced by luminal injection of toxin A into the mouse gut. Mechanistically, we found that Peri-4 could reduce toxin A-induced reactive oxygen species production to inhibit the activations of p38MAPK and p21Cip1/Waf1 , which are critical for the cell damages induced by toxin A. These results collectively suggest that the Peri-4 may be a potential therapeutic agent for treating toxin A-induced pseudomembranous colitis. Copyright © 2017 European Peptide Society and John Wiley & Sons, Ltd.

Expression patterns of members of the ethylene signaling-related gene families in response to dehydration stresses in cassava.

Drought is the one of the most important environment stresses that restricts crop yield worldwide. Cassava (Manihot esculenta Crantz) is an important food and energy crop that has many desirable traits such as drought, heat and low nutrients tolerance. However, the mechanisms underlying drought tolerance in cassava are unclear. Ethylene signaling pathway, from the upstream receptors to the downstream transcription factors, plays important roles in environmental stress responses during plant growth and development. In this study, we used bioinformatics approaches to identify and characterize candidate Manihot esculenta ethylene receptor genes and transcription factor genes. Using computational methods, we localized these genes on cassava chromosomes, constructed phylogenetic trees and identified stress-responsive cis-elements within their 5' upstream regions. Additionally, we measured the trehalose and proline contents in cassava fresh leaves after drought, osmotic, and salt stress treatments, and then it was found that the regulation patterns of contents of proline and trehalose in response to various dehydration stresses were differential, or even the opposite, which shows that plant may take different coping strategies to deal with different stresses, when stresses come. Furthermore, expression profiles of these genes in different organs and tissues under non-stress and abiotic stress were investigated through quantitative real-time PCR (qRT-PCR) analyses in cassava. Expression profiles exhibited clear differences among different tissues under non-stress and various dehydration stress conditions. We found that the leaf and tuberous root tissues had the greatest and least responses, respectively, to drought stress through the ethylene signaling pathway in cassava. Moreover, tuber and root tissues had the greatest and least reponses to osmotic and salt stresses through ethylene signaling in cassava, respectively. These results show that these plant tissues had differential expression levels of genes involved in ethylene signaling in response to the stresses tested. Moreover, after several gene duplication events, the spatiotemporally differential expression pattern of homologous genes in response to abiotic and biotic stresses may imply their functional diversity as a mechanism for adapting to the environment. Our data provide a framework for further research on the molecular mechanisms of cassava resistance to drought stress and provide a foundation for breeding drought-resistant new cultivars.

NQO1-Knockout Mice Are Highly Sensitive to Clostridium Difficile Toxin A-Induced Enteritis.

Clostridium difficile toxin A causes acute gut inflammation in animals and humans. It is known to downregulate the tight junctions between colonic epithelial cells, allowing luminal contents to access body tissues and trigger acute immune responses. However, it is not yet known whether this loss of the barrier function is a critical factor in the progression of toxin A-induced pseudomembranous colitis. We previously showed that NADH:quinone oxidoreductase 1 (NQO1) KO (knockout) mice spontaneously display weak gut inflammation and a marked loss of colonic epithelial tight junctions. Moreover, NQO1 KO mice exhibited highly increased inflammatory responses compared with NQO1 WT (wild-type) control mice when subjected to DSS-induced experimental colitis. Here, we tested whether toxin A could also trigger more severe inflammatory responses in NQO1 KO mice compared with NQO1 WT mice. Indeed, our results show that C. difficile toxin A-mediated enteritis is significantly enhanced in NQO1 KO mice compared with NQO1 WT mice. The levels of fluid secretion, villus disruption, and epithelial cell apoptosis were also higher in toxin A-treated NQO1 KO mice compared with WT mice. The previous and present results collectively show that NQO1 is involved in the formation of tight junctions in the small intestine, and that defects in NQO1 enhance C. difficile toxin A-induced acute inflammatory responses, presumably via the loss of epithelial cell tight junctions.

Potassium Acetate Blocks Clostridium difficile Toxin A-Induced Microtubule Disassembly by Directly Inhibiting Histone Deacetylase 6, Thereby Ameliorating Inflammatory Responses in the Gut.

Clostridium difficile toxin A is known to cause deacetylation of tubulin proteins, which blocks microtubule formation and triggers barrier dysfunction in the gut. Based on our previous finding that the Clostridium difficile toxin A-dependent activation of histone deacetylase 6 (HDAC-6) is responsible for tubulin deacetylation and subsequent microtubule disassembly, we herein examined the possible effect of potassium acetate (PA; whose acetyl group prevents the binding of tubulin to HDAC-6) as a competitive/false substrate. Our results revealed that PA inhibited toxin A-induced deacetylation of tubulin and recovered toxin A-induced microtubule disassembly. In addition, PA treatment significantly decreased the production of IL-6 (a marker of inflamed tissue) in the toxin A-induced mouse enteritis model. An in vitro HDAC assay revealed that PA directly inhibited HDAC-6-mediated tubulin deacetylation, indicating that PA acted as a false substrate for HDAC-6. These results collectively indicate that PA treatment inhibits HDAC-6, thereby reducing the cytotoxicity and inflammatory responses caused by C. difficile toxin A.

The Insect Peptide CopA3 Increases Colonic Epithelial Cell Proliferation and Mucosal Barrier Function to Prevent Inflammatory Responses in the Gut.

The epithelial cells of the gut form a physical barrier against the luminal contents. The collapse of this barrier causes inflammation, and its therapeutic restoration can protect the gut against inflammation. EGF enhances mucosal barrier function and increases colonocyte proliferation, thereby ameliorating inflammatory responses in the gut. Based on our previous finding that the insect peptide CopA3 promotes neuronal growth, we herein tested whether CopA3 could increase the cell proliferation of colonocytes, enhance mucosal barrier function, and ameliorate gut inflammation. Our results revealed that CopA3 significantly increased epithelial cell proliferation in mouse colonic crypts and also enhanced colonic epithelial barrier function. Moreover, CopA3 treatment ameliorated Clostridium difficile toxin As-induced inflammation responses in the mouse small intestine (acute enteritis) and completely blocked inflammatory responses and subsequent lethality in the dextran sulfate sodium-induced mouse model of chronic colitis. The marked CopA3-induced increase of colonocyte proliferation was found to require rapid protein degradation of p21(Cip1/Waf1), and an in vitro ubiquitination assay revealed that CopA3 directly facilitated ubiquitin ligase activity against p21(Cip1/Waf1). Taken together, our findings indicate that the insect peptide CopA3 prevents gut inflammation by increasing epithelial cell proliferation and mucosal barrier function.

Antimicrobial Peptide, Lumbricusin, Ameliorates Motor Dysfunction and Dopaminergic Neurodegeneration in a Mouse Model of Parkinson's Disease.

We recently reported that the antimicrobial peptide Lumbricusin (NH2-RNRRWCIDQQA), isolated from the earthworm, increases cell proliferation in neuroblastoma SH-SY5Y cells. Here, we investigated whether Lumbricusin has neurotropic activity in mouse neural stem cells (MNSCs) and a protective effect in a mouse model of Parkinson's disease (PD). In MNSCs isolated from mouse brains, Lumbricusin treatment significantly increased cell proliferation (up to 12%) and reduced the protein expression of p27(Kip1) through proteasomal protein degradation but not transcriptional regulation. Lumbricusin inhibited the 6-OHDA-induced apoptosis of MNSCs, and also showed neuroprotective effects in a mouse PD model, ameliorating the motor impairments seen in the pole, elevated body swing, and rotation tests. These results suggest that the Lumbricusin-induced promotion of neural cell proliferation via p27(Kip1) degradation has a protective effect in an experimental PD model. Thus, the antimicrobial peptide Lumbricusin could possibly be developed as a potential therapeutic agent for the treatment of PD.

[Characteristics of 72 h glucose profiles detected by continuous glucose monitoring system in patients with insulinoma].

OBJECTIVE: To identify the characteristics of glucose profiles in patients with insulinoma using continuous glucose monitoring system (CGMS). METHODS: Six patients with insulinoma (All of the patients were diagnosed after operations with pathological tests) admitted to the Department of Endocrinology and Metabolism, West China Hospital of Sichuan University from October 2009 to November 2011 were recruited for this study (group A). They were compared with 71 patients with normal glucose tolerance (NGT) (group B) and 3 patients with responsive hypoglycemia (group C). All of the participants received 75 g oral glucose tolerance tests (OGTT), insulin release tests, and 72 h CGM. Glucose fluctuations and hypoglycemia risks were assessed by the following parameters obtained from CGMS: mean blood glucose (MBG) and standard deviation (SD), mean amplitude of glycemic excursion (MAGE), mean daily differences (MODD), low glucose index (LBGI), glycaemic Risk Assessment Diabetes Equation (GRADE) and M-value. RESULTS: Group A had significantly lower levels of HbA1c, fasting blood glucose, and 2 h-BG than group B (P < 0.05). Compared with Group B, Group A had lower levels of 72 h-MBG (P < 0.001) and higher levels of LBGI, M-value, GRADE, SD and MODD (P < 0.05). The insulinoma patients had 27 occasions of hypoglycemia (glucose < 2.8 mmol/L), more likely in early morning, at a fasting state or preprandial periods. Such events were rare in the control groups (2 occasions in Group B and one occasion in Group C). CONCLUSION: Insulinoma patients present a greater level of glucose fluctuations and frequent hypoglycemia. Hypoglycemia risk detected by CGM is helpful for differentiating diagnosis of insulinoma.

[Effect of mixed interface on the performance of solution-processed phosphorescent OLEDs].

In the present work, in order to improve electron injection and transport at the interface of the hole blocking layer (HBL) and the electron transport layer (ETL) in the hole-domain solution processed phosphorescent organic light emitting devices (PhOLEDs), the mixed interface layer (MIL) was fabricated by partially co-doping hole blocking material 2,9-dimethyl-4,7-diphenyl-1,10-phenanthroline (BCP) and electron transport material tris(8-quinolinolato) aluminum (Alq3) between HBL/ ETL. The MIL thickness was kept at 10nm, while the doping ratio of these two materials varied. Under a given electric field, the devices with the MIL at any mixed ratios all show much higher luminance and current density than those with a typical interface. For example, the luminance power and current density at 10 V for a typical device are 1.03 microW and 5.13 mA x cm(-2), while in case of mixed interface are 3.64 microW and 18.1 mA x cm(-2), respectively. From data results and theoretical analysis, the possible derivation of these improvements is considered to be the reduced electron accumulation at the interface resulting from the reduced electron injection energy barrier and lowered transport mobility by BCP material, which leads to an increase in electron amount in the emission layer and therefore the higher luminance and current density.

[Estimating intraday blood glucose variability through self-monitoring of blood glucose in people with impaired glucose tolerance and type 2 diabetes].

OBJECTIVE: To investigate the associations between the patterns of change of self-monitored blood glucose (SMBG) and the parameters of intraday blood glucose variability [mean absolute glucose excursions (MAGE), mean postprandial glucose excursion (MPPGE) and standard deviation of blood glucose (sBG)] measured by the continuous glucose monitoring system. METHODS: A 72-hour continuous glucose monitoring was performed in a sample 105 people with impaired glucose tolerance (IGR, n=51) and newly-diagnosed type 2 diabetes mellitus (T2DM, n=54) to calculate MAGE, MPPGE and sBG. Meanwhile, fingertip blood glucose self-monitoring was performed to determine fasting blood glucose (FBG), blood glucose after breakfast (BG(AB)), blood glucose before lunch (BG(BL)), blood glucose after lunch (BG(AL)), blood glucose before supper (BG(BS)), blood glucose after supper (BG(AS)), and blood glucose before sleeping (BG(BR)) at the same period of time. Multiple stepwise regression analysis was performed to generate equations for predicting MAGE, MPPGE and sBG with age and the self-monitoring blood glucose parameters in 80% of the subjects (41 IGR and 44 T2DM, randomly selected from the overall sample). These equations were then cross-validated in the remaining 20% subjects (10 IGR and 10 T2DM). RESULTS: BG(AA), BG(AB), BGAL and FBG entered into the regression equations predicting MAGE, sBG and MPPGE for the IGR subject, while age only entered into the regression equations predicting MPPGE and sBG. For the subjects with T2DM, BG(AS), BG(AL) and age entered into the equation predicting MAGE; BG(AS), BG(AL), BG(BL) and BG(BS) entered into the equation predicting MPPGE; BG(AS), BG(AL) and FBG entered into the equation predicting sBG. The cross-validation study showed that the differences between predicted and observed values of MAGE in the subjects with IGR and T2DM were 4.1% and 8.2%, respectively; the differences between predicted and observed values of MPPGE in the subjects with IGR and T2DM were 23.1% and 1.3%, respectively; and the differences between predicted and observed values of sBG in the subjects with IGR and T2DM were 1.2% and 6.8%, respectively. Except for MPPGE in the subjects with IGR, the goodness of fit between predicted and observed values were good. CONCLUSION: The MAGE and sBG in people with IGR and the MAGE, MPPGE and sBG in patients with T2DM can be well predicted with age and self-monitored blood glucose.

[Preparation and transmissivity of ZnS nanocolumn thin films with glancing angle deposition technology].

Nanocrystalline ZnS thin films were fabricated by glancing angle deposition (GLAD) technology in an electron beam evaporation system. Deposition was carried out in the custom vacuum chamber at a base pressure 3 x 10(-4) Pa, and the deposition rate was fixed at 0.2 nm x s(-1). ZnS films were deposited on pieces of indium tin oxide (ITO) substrates when the oblique angle of the substrate relative to the incoming molecular flux was set to 0 degrees, 80 degrees and 85 degrees off the substrate normal respectively. X-ray diffraction (XRD) spectra and scanning electron microscope (SEM) images showed that ZnS nanocrystalline films were formed on the substrates at different oblique angle, but the nanocolumn structure was only formed under the situation of alpha = 80 degrees and 85 degrees. The dynamics during the deposition process of the ZnS films at alpha = 0 degrees, 80 degrees and 85 degrees was analyzed. The transmitted spectra of ZnS thin films deposited on ITO substrates showed that the ZnS nanocolumn thin films could enhance the transmissivity in visible range. The ZnS nanocolumn could be used into electroluminescence device, and it would enhance the luminous efficiency of the device.

[Accuracy and safety of continuous glucose monitoring system in diabetic and non-diabetic subjects].

OBJECTIVE: To evaluate the accuracy and safety of continuous glucose monitoring system (CGMS) in subjects with normal glucose tolerance (NGT), impaired glucose regulation (IGR) and newly-diagnosed type 2 diabetes mellitus (T2DM). METHODS: A total of 162 subjects (53 NGT, 53 IGR and 56 newly diagnosed T2DM) at 22 - 72 years old were recruited. Each subject received a continuous glucose monitoring (CGM) of CGMS SYSTEM GOLD(TM) (Medtronic Minimed, Northridge, CA) for 3 consecutive days and was instructed to self-calibrate the interstitial glucose levels with finger-stick blood glucose measurements (self-monitoring of blood glucose, SMBG) 7 times daily. Means of median absolute difference (median AD) and median absolute relative difference (median RAD) were calculated to assess the difference between CGM and SMBG values. The correlation between CGM and SMBG values were analyzed with the liner regression analysis. The data were analyzed by the ISO criteria for home glucose meters and Clarke error grid analysis. RESULTS: All participants showed a good tolerance to the insertion of CGMS sensor and wearing the device. The mean duration of CGMS recording was (75.6 ± 8.3) h. A total of 4324 glucose meter values were paired with glucose measurements from CGMS. Overall, a good relationship (r = 0.943, P = 0.000) and no significant difference (P > 0.05) over a wide range (2.2 - 22.2 mmol/L) were found between CGM and SMBG values. The median AD was 0.5 (0.2 - 0.9) mmol/L and the median RAD 7.55% (3.33% - 14.09%) with 87.16% of sensor values meeting the ISO home glucose meter criteria. The Clarke error grid analysis showed that 99.58% of the readings from CGMS fell into the clinical acceptable zones including 87.5% values in zone A (clinical exact) and 12.1% values in zone B (benign error). CONCLUSION: Both safe and well-tolerated, continuous glucose monitoring provides comparatively accurate blood glucose values to guide the diagnosis and treatment of diabetes.

Stabilization of recombinant human growth hormone against emulsification-induced aggregation by Pluronic surfactants during microencapsulation.

Protein aggregation upon exposing to the water/organic solvent interface is one of the most significant obstacles in developing poly(lactic-co-glycolic acid) (PLGA) microspheres with double emulsion process. The aim of present study is to devise a formulation strategy to prevent recombinant human growth hormone (rhGH) from aggregation during microencapsulation. The excipients used for stabilizing rhGH were selected from sugars, nonionic surfactants, polyol, and protein. Among the candidates, surfactants exhibited potentialities in protecting rhGH against emulsification-induced aggregation. It was also found that Pluronic F127 showed an outstanding as well as concentration-dependent stabilizing effect on rhGH, which was different to Pluronic F68 and Tween 20. After the rhGH solution comprising F127 and sucrose was emulsified with methylene chloride, the recovery of monomeric protein achieved 99.0%, principally attributed to the presence of F127. This solution was subsequently encapsulated as inner aqueous phase in the PLGA microspheres by a conventional double emulsion process, with the encapsulation efficiency higher than 98%. Improvement in the release of rhGH was observed for the microspheres co-encapsulating Pluronic F127 regardless in the presence or absence of sucrose, compared to the microspheres containing rhGH alone. The result further implied that co-encapsulation of Pluronic F127 in the microspheres played an important role in the stabilization of rhGH.