Silencing of miR-330-5p stimulates osteogenesis in bone marrow mesenchymal stem cells and inhibits bone loss in osteoporosis by activating Bgn-mediated BMP/Smad pathway.

OBJECTIVE: To illustrate the role of micro ribonucleic acid (miR)-330-5p in regulating osteogenesis through biglycan (Bgn)-mediated bone morphogenetic protein (BMP)/Smad pathway. MATERIALS AND METHODS: A mouse model of osteoporosis (OP) was established by ovariectomy (OVX). BMD and miR-330-5p levels in mice undergoing sham operation or OVX were determined. BMD and BV/TV in OP mice with in vivo knockdown of miR-330-5p were measured by Micro-CT. After silencing of miR-330-5p in mouse primary bone marrow stromal cells (BMSCs), expression changes in osteogenesis-associated genes, ALP activity, and mineralization ability were assessed. Subsequently, the interaction between miR-330-5p and Bgn was examined by Dual-Luciferase reporter gene assay and Western blotting. Then, Bgn levels in BMSCs undergoing osteogenesis at different time points were measured. At last, the regulatory effects of miR-330-5p/Bgn axis on the BMP/Smad pathway, ALP activity, and mineralization ability in BMSCs were evaluated. RESULTS: BMD was decreased and miR-330-5p was upregulated in OP mice. OP mice with in vivo knockdown of miNA-330-5p presented higher BMD and BV/TV than controls. Transfection with miR-330-5p inhibitor upregulated osteogenesis-associated genes, ALP activity, and mineralization ability in BMSCs. Bgn was time-dependently upregulated in BMSCs undergoing osteogenesis, which was indicated to be the target gene of miR-330-5p. Besides, Bgn level was negatively regulated by miR-330-5p. Importantly, Bgn was able to reverse the regulatory effects of miR-330-5p on the BMP/Smad pathway, ALP activity, and mineralization ability in BMSCs. CONCLUSIONS: Knockdown of miR-330-5p facilitates osteogenesis in BMSCs through the Bgn-induced BMP/Smad pathway, thus alleviating the progression of OP.

Solar PV-Battery-Electric Grid-Based Energy System for Residential Applications: System Configuration and Viability.

Distributed renewable energy share increase in electricity generation is creating challenges for the whole power system, due to its intermittent and nonprogrammable nature. Energy storage has the potential to solve those issues although its technical, economic, and environmental impact is up for debate. The paper presents a study about a PV-battery energy storage system installed in a grid-connected residential apartment in the Green Energy Laboratory at Shanghai Jiao Tong University, China. Daily experimental results show how the presence of energy storage reduces the midday feed-in of excess PV power and the evening peak demand, providing benefits to the distribution network in terms of reduced voltage swings and peak load. Considering the Chinese context, an economic analysis is carried out to assess the profitability of residential PV-battery systems, using the net present value as the economic indicator of an 18-year investment in which the battery pack is replaced twice (6 life years). The analysis shows that such system is not economically viable due to a combination of low electricity prices, valuable PV incentives, and high technology costs. However, considering a future scenario of doubled electricity tariff, halved export tariff, and falling technology costs (-66% battery and -17% PV and inverter), PV-battery investment becomes profitable and shows more resilience to future scenarios than PV-only investment.

High non-esterified fatty acid concentrations promote expression and secretion of fibroblast growth factor 21 in calf hepatocytes cultured in vitro.

Negative energy balance is considered as the pathological basis of energy metabolic disorders in periparturient dairy cows. Serum non-esterified fatty acids (NEFA) are one of the most important indicators of energy balance status. Fibroblast growth factor 21 (FGF21) has been identified as a hepatokine involved in regulation of metabolic adaptations, such as promoting hepatic lipid oxidation and ketogenesis, during energy deprivation. However, the direct effects of NEFA on FGF21 expression and secretion in bovine hepatocytes are not entirely clear. The objective of this study was to investigate the effects of different NEFA concentrations on FGF21 expression and secretion in calf hepatocytes cultured in vitro. NEFA were added to the culture solution at final concentrations of 0.6, 1.2, 1.8 and 2.4 mmol/L. After 24 hr of continuous culture, FGF21 mRNA and protein expression levels in the hepatocytes were determined by real-time PCR and Western blot respectively. FGF21 secretion in the supernatant was determined by enzyme-linked immunosorbent assay (ELISA). The results showed that expression and secretion of FGF21 at 0.6 mmol/L NEFA-treated hepatocytes was higher than that of the control group (p < .05). The FGF21 expression and secretion were similar at 1.2, 1.8 and 2.4 mmol/L NEFA-treated hepatocytes and significantly higher than those observed for controls (p < .01). These data suggest that high concentrations of NEFA significantly promote FGF21 expression and secretion in bovine hepatocytes. In particular, this promotion occurs in a dose-dependent manner and may be involved in the pathological processes of energy metabolism disorders of dairy cows in the peripartum period.

[Autologous ipsilateral lamellar corneal-flap transposition for treatment of peripheral corneal perforation].

OBJECTIVE: To study the clinical outcome of autologous ipsilateral lamellar corneal-flap transposition for treatment of peripheral corneal perforation. METHODS: Medical records of 13 patients who underwent autologous ipsilateral lamellar corneal-flap transposition for peripheral corneal perforation from January 2008 to February 2015 were retrospectively reviewed. The diameter of the corneal ulcers was all ≤ 3.0 mm, and the diameter of perforations was all ≤ 2.0 mm. All patients were followed up for 3-12 months with an average of 8 months. The healing of ulcer, clarity of the grafts, pre-and postoperative visual acuity, and astigmatism were documented and analyzed. RESULTS: At 1 day after surgery, all patients had normal anterior chamber depth without aqueous leak. No double chamber or iris synechia was found. The corneal grafts showed mild to moderate edema. On postoperative days 7-14, the corneal edema receded, the grafts became transparent gradually, and the ulcer margin was blurred. Three months after surgery, all the grafts remained transparent. There was no recurrence of ulcer during the follow-up period. The best corrected visual acuity before and after surgery was 0.44 ± 0.19 and 0.47 ± 0.20, respectively, which indicated no statistically significant difference(t=0.706, P=0.461). The corneal astigmatism before and after surgery was 0.44±0.19 and 0.47±0.20, respectively, which showed statistically significant difference(t=2.391, P<0.05). CONCLUSIONS: Autologous ipsilateral lamellar corneal-flap transposition is a safe and effective surgical procedure for treating peripheral corneal perforation. It can be considered as an alternative method for penetrating or lamellar keratoplaty when the availability of corneal tissue is limited, or for cases with a high risk of developing immunologic allograft rejection, although the surgical indications are limited. (Chin J Ophthalmol, 2016, 52: 663-668).

LaPtSb: a half-Heusler compound with high thermoelectric performance.

The electronic and transport properties of the half-Heusler compound LaPtSb are investigated by performing first-principles calculations combined with semi-classical Boltzmann theory and deformation potential theory. Compared with many typical half-Heusler compounds, LaPtSb exhibits an obviously larger power factor at room temperature, especially for the n-type system. Together with the very low lattice thermal conductivity, the thermoelectric figure of merit (ZT) of LaPtSb can be optimized to a record high value of 2.2 by fine tuning the carrier concentration.

Uniformly carbon-covered alumina and its surface characteristics.

Uniformly carbon-covered alumina (CCA) was prepared via the carbonization of sucrose highly dispersed on the alumina surface. The CCA samples were characterized by XRD, XPS, DTA-TG, UV Raman, nitrogen adsorption experiments at 77 K, and rhodamine B (RB) adsorption in aqueous media. UV Raman spectra indicated that the carbon species formed were probably conjugated olefinic or polycyclic aromatic hydrocarbons, which can be considered molecular subunits of a graphitic plane. The N(2) adsorption isotherms, pore size distributions, and XPS results indicated that carbon was uniformly dispersed on the alumina surface in the as-prepared CCA. The carbon coverage and number of carbon layers in CCA could be controlled by the tuning of the sucrose content in the precursor and impregnation times. RB adsorption isotherms suggested that the monolayer adsorption capacity of RB on alumina increased drastically for the sample with uniformly dispersed carbon. The as-prepared CCA possessed the texture of alumina and the surface properties of carbon or both carbon and alumina depending on the carbon coverage.

The Rxo1/ Rba1 locus of maize controls resistance reactions to pathogenic and non-host bacteria.

Infiltration of different maize lines with a variety of bacterial pathogens of maize, rice and sorghum identified qualitative differences in resistant reactions. Isolates from two bacterial species induced rapid hypersensitive reactions (HR) in some maize lines, but not others. All isolates of the non-host pathogen Xanthomonas oryzae pv. oryzicola (bacterial leaf streak disease of rice) and some isolates of the pathogenic bacterium Burkholderia andropogonis induced HR when infiltrated into maize line B73, but not Mo17. Genetic control of the HR to both bacteria segregated as a single dominant gene. Surprisingly, both phenotypes mapped to the same locus, indicating they are either tightly linked or controlled by the same gene. The locus maps on the short arm of maize chromosome six near several other disease-resistance genes. Results indicate the same type of genes may contribute to both non-host resistance and resistance to pathogens.

Differentiation of monocytes to macrophages switches the Mycobacterium tuberculosis effect on HIV-1 replication from stimulation to inhibition: modulation of interferon response and CCAAT/enhancer binding protein beta expression.

HIV-1 replication is inhibited in uninflamed lung macrophages and is stimulated during tuberculosis. Attempts to recapitulate activation of HIV-1 replication in primary monocytes and macrophages ex vivo and in the untreated and PMA-treated THP-1 cell line model in vitro have produced opposite results depending on the state of differentiation of the cells. After infection with Mycobacterium tuberculosis, monocytes enhanced HIV-1 replication and produced a stimulatory 37-kDa CCAAT/enhancer binding protein beta (C/EBPbeta) transcription factor, whereas macrophages suppressed HIV-1 replication and produced an inhibitory 16-kDa C/EBPbeta transcription factor. IFN-beta induced inhibitory 16-kDa C/EBPbeta in macrophages, but had no effect on C/EBPbeta expression in monocytes. Macrophages, but not monocytes, were able to activate IFN-stimulated gene factor-3 (ISGF-3), a transcription factor composed of STAT-1, STAT-2, and IFN regulatory factor (IRF)-9, after infection with M. tuberculosis or stimulation with type I IFN. Macrophages expressed IRF-9 DNA-binding activity, but monocytes did not, and addition of the IRF-9 component reconstituted ISGF-3 in extracts of IFN-treated monocytes. Modulation of IFN responsiveness upon differentiation occurred at least in part through a post-transcriptionally regulated increase in IRF-9 expression. Both monocytes and macrophages maintained IFN responsiveness, activating STAT-1 homodimer formation and transcription of the STAT-1 gene after IFN stimulation. In addition, both monocytes and macrophages were able to activate NF-kappaB upon infection with M. tuberculosis. These results show that induction of ISGF-3, expression of the inhibitory 16-kDa C/EBPbeta, and suppression of HIV-1 replication via a transcriptional mechanism are macrophage-specific responses to infection with M. tuberculosis.

[Effect of corticotrophin on formaldehyde-induced somatostatin of spinal cord in rats].

AIM: To study the effect of corticotrophin (Cor) on somatostatin (Som) and its synthesis in spinal dorsal horn induced by formaldehyde in rats. METHODS: Using double immunohistochemical stainings, and in situ hybridization. RESULTS: Two hours after s.c. formaldehyde (5%, 200 microL) in one hindpaw of rats, the neurons of c-fos-like immunoreactivity (FLI), somatostatin-like immunoreactivity (Som-LI), Som-LI/FLI, and perprosomatostatin mRNA (PPS-mRNA) in ipsilateral spinal dorsal horn were increased obviously, as compared with the control group. The FLI and Som-LI of spinal cord were not changed by i.p. Cor. Cor (25 or 12.5 U.kg-1, i.p.) inhibited the formaldehyde-evoked FLI, Som-LI, Som-LI/FLI, and PPS-mRNA of spinal cord in a dose-dependent manner. The decrease of c-fos or Som level due to i.p. Cor in rats of chronic pain was prevented by raphe nuclei injected cyproheptadine, but not by bicuculline, naloxone, or phentolamin injected to raphe nuclei. CONCLUSION: The formaldehyde-evoked c-fos expression, Som, and Som synthesis of spinal cord were suppressed by Cor through the serotonin receptor of raphe nuclei.

Fluoroquinolone action against clinical isolates of Mycobacterium tuberculosis: effects of a C-8 methoxyl group on survival in liquid media and in human macrophages.

When the lethal action of a C-8 methoxyl fluoroquinolone against clinical isolates of Mycobacterium tuberculosis in liquid medium was measured, the compound was found to be three to four times more effective (as determined by measuring the 90% lethal dose) than a C-8-H control fluoroquinolone or ciprofloxacin against cells having a wild-type gyrA (gyrase) gene. Against ciprofloxacin-resistant strains, the C-8 methoxyl group enhanced lethality when alanine was replaced by valine at position 90 of the GyrA protein or when aspartic acid 94 was replaced by glycine, histidine, or tyrosine. During infection of a human macrophage model by wild-type Mycobacterium bovis BCG, the C-8 methoxyl group lowered survival 20- to 100-fold compared with the same concentration of a C-8-H fluoroquinolone. The C-8 methoxyl fluoroquinolone was also more effective than ciprofloxacin against a gyrA Asn94 mutant of M. bovis BCG. In an M. tuberculosis-macrophage system the C-8 methoxyl group improved fluoroquinolone action against both quinolone-susceptible and quinolone-resistant clinical isolates. Thus, a C-8 methoxyl group enhances the bactericidal activity of quinolones with N1-cyclopropyl substitutions; these data encourage further refinement of fluoroquinolones as antituberculosis agents.

Intracellular fate of Mycobacterium avium subspecies paratuberculosis in monocytes from normal and infected, interferon-responsive cows as determined by a radiometric method.

The ability of Mycobacterium avium subsp. paratuberculosis to survive in bovine monocytes was studied using radiometric (BACTEC) culture, standard plate counting and microscopic counting of acid-fast stained monocyte monolayers. Results of microscopic counts sharply contrasted with results of viable counts determined both by plate counting and radiometric counting. We observed an early phase (the first 6 d after in vitro infection) of intracellular bacillary growth, followed by a later phase of mycobacteriostasis or killing (up to 12 d after in vitro infection) in monocytes from non-infected cows. The data suggest that multiplication and death of M. avium subsp. paratuberculosis occur simultaneously in bovine monocytes infected in vitro. Using the BACTEC method, we compared the ability of bovine monocytes from normal cows and cows infected with M. avium subsp. paratuberculosis and showing evidence of a strong Thl-like cellular immune response to ingest and inhibit the intracellular growth of M. avium subsp. paratuberculosis. There was a trend toward greater phagocytosis and faster killing of Mycobacterium avium subsp. paratuberculosis by monocytes from the infected, immune responder cows. However, the observed numbers of viable M. avium subsp. paratuberculosis at each time after monocyte infection were not significantly different between normal and infected cows.

Intrathecal injection of corticotropin inhibited nitric-oxide synthase-positive neuron increase in rat spinal cord after formalin-induced hyperalgesia.

AIM: To study the effects of corticotropin (Cor) on formalin-induced hyperalgesia and the change of nitric-oxide synthase (NOS)-positive neurons in spinal dorsal horn in rats. METHODS: Measurement of pain intensity rating (PIR), NADPH-d histochemistry, and Fos immunohistochemistry were adopted. RESULTS: The increases of NOS-positive neurons, Fos, NOS/Fos double labelling neurons of the spinal dorsal horn and the PIR after formalin injection were markedly inhibited by intrathecal injecting (ith) Cor (0.5-1.5 U), which were obviously attenuated by L-arginine (Arg, 5-15 nmol, ith), the substrate of NOS. CONCLUSION: Cor inhibits formalin-induced hyperalgesia by the decrease of NOS-positive neurons in the spinal dorsal horn of rats.

Type I interferon induces inhibitory 16-kD CCAAT/ enhancer binding protein (C/EBP)beta, repressing the HIV-1 long terminal repeat in macrophages: pulmonary tuberculosis alters C/EBP expression, enhancing HIV-1 replication.

We have previously observed that HIV-1 replication is suppressed in uninflamed lung and increased during tuberculosis. In vitro THP-1 cell-derived macrophages inhibited HIV-1 replication after infection with Mycobacterium tuberculosis. Suppression of HIV-1 replication was associated with inhibition of the HIV-1 long terminal repeat (LTR) and induction of ISGF-3, a type I interferon (IFN)-specific transcription factor. Repression of the HIV-1 LTR required intact CCAAT/enhancer binding protein (C/EBP) sites. THP-1 cell-derived macrophages infected with M. tuberculosis, lipopolysaccharide, or IFN-beta induced the 16-kD inhibitory C/EBPbeta isoform and coincidentally repressed HIV-1 LTR transcription. C/EBPbeta was the predominant C/EBP family member produced in THP-1 macrophages during HIV-1 LTR repression. In vivo, alveolar macrophages from uninflamed lung strongly expressed inhibitory 16-kD C/EBPbeta, but pulmonary tuberculosis abolished inhibitory C/EBPbeta expression and induced a novel C/EBP DNA binding protein. Therefore, in vitro, proinflammatory stimulation produces an IFN response inhibiting viral replication by induction of a C/EBPbeta transcriptional repressor. THP-1 cell-derived macrophages stimulated with type I IFN are similar to alveolar macrophages in the uninflamed lung in vivo. In contrast, the cellular immune response in active pulmonary tuberculosis disrupts this innate immunity, switching C/EBP expression and allowing high level viral replication.

[Somatostatin and electroacupuncture inhibited c-fos expression in spinal cord of arthritic rats].

AIM: To observer the effect of intrathecal injection of somatostatin (Som) associated with electroacupuncture (EA) at "Jiaji" points on c-fos protein expression of spinal cord in pain rats. METHODS: Rats with adjuvant arthritis were used as pain model and the c-fos-like immunoreactivity (FLI) was detected by immunohistochemistry. RESULTS: The c-fos protein expression induced by arthritis were found in all of the I-X laminae of ipsilateral spinal cord of rats, and most of the labeled cells were located in the laminae I-II and V-VI. Som and EA suppressed the c-fos expression and the lessening of FLI cells in the spinal cord. CONCLUSION: Pathological pain following arthritis activated pain sensitive neurons (PSN) and evoked c-fos expression in spinal cord, Som and EA suppressed activities of these PSN, producing the effect of analgesia.

Induction of L-arginine-dependent production of nitric oxide in bovine monocytes by interferon gamma and lipopolysaccharide.

Bovine monocytes freshly isolated from peripheral blood were induced to produce nitric oxide by exposing them to recombinant bovine interferon gamma (rbIFN-y) and Escherichia coli lipopolysaccharide (LPS) in vitro. Moderate amounts of nitric oxide were induced by rbIFN-gamma alone, but larger amounts were induced by rbIFN-gamma and LPS together, the amount being dependent on the quantity of rbIFN-gamma added. Reactive nitrogen intermediates (RNI) were produced within six hours, their concentration peaked at four days and they were detectable for at least eight days after the cells had been stimulated with rbIFN-gamma and LPS. The production of RNI was diminished by the addition of NG-monomethyl-L-arginine. The data suggest that bovine monocytes can produce RNI via a pathway involving an inducible nitric oxide synthase.

Arboviruses as aetiological agents of encephalitis in the People's Republic of China.

A serological study was undertaken to determine the role of arboviruses as etiological agents of encephalitis in the People's Republic of China (PRC). Paired sera were collected during mosquito seasons in 1988-1990 from 614 patients with possible viral encephalitis in 15 regions of PRC and tested for haemagglutination inhibiting antibodies to selected arboviruses. Seroconversions were documented to alphavirus and flavivirus antigens in 13.0 and 18.7% of patients respectively in most of the study areas. No California group seroconversion was detected. The age of alphavirus seroconvertors ranged from 2 months to 32 years and of flavivirus seroconvertors from 6 months to 50 years, with higher numbers in males. Serious central nervous system manifestations were seen more commonly in flavivirus seroconvertors. This study affirms the importance of flavivirus as causative agents of encephalitis in PRC and provides evidence that one or more alphaviruses are causing symptomatic infections with neurological involvement in PRC.

[Changes of the 5-HT content in hippocampus, midbrain-pons, spinal cord and concentration of blood glucose after intraperitoneal injection of ACTH in rats].

Effects of ACTH on the content of 5-HT in the hippocampus, midbrain-pons, and spinal cord and the concentration of blood glucose in rats by spectrofluorometric assay and glucose oxidase method were studied. ACTH ip 10 IU.kg-1 or 20 IU.kg-1 significantly increased the content of 5-HT in the hippocampus, midbrain-pons and spinal cord and blood glucose level, both in a dose-dependent manner; para-chloroaphetamine, p-Cpa 4 mg icv markedly reduced the 5-HT in these brain regions and spinal cord and the blood glucose level (P less than 0.01); the level of 5-HT and blood glucose were not significantly altered after icv p-Cpa+ip ACTH; adrenalectomy+ip ACTH markedly increased the 5-HT content in the brain regions and spinal cord, but the blood glucose was decreased (P less than 0.01); and 5-HT was markedly decreased by sc alloxan tetrahydrate+ip ACTH, but blood glucose did not decrease (P greater than 0.05). Thus, ACTH may influence the blood glucose level through central nervous system 5-HT, and the change of 5-HT may be related to the insulin (secretion).

[Effect of intraventricular injection of somatostatin on pain threshold, and contents of the monoamines, xanthine, hypoxanthine in rats brain].

Using HPLC with electrochemical detection, we found that icv somatostatin (Som) 5 or 10 micrograms increased rat's pain threshold and contents of 5-HT and 5-HIAA in hippocampus, hypothalamus and brainstem, except the 5-HIAA content of brainstem in Som 5 micrograms group. However, the changes of NE among above three areas of brain were different, the NE contents of hypothalamus and brainstem significantly increased while that of hippocampus markedly decreased. After icv Som 20 micrograms, hypoxanthine and xanthine in hippocampus and hypothalamus decreased significantly, but encephaledema occurred. Som 40 micrograms icv caused necrotic changes of neurons in brain.

[Serotonin of hippocampus and hypothalamus taking part in the analgesic effect of adrenocorticotropic hormone in rats].

The effects of ip intra-PAG injection of ACTH on serotonin (5-HT), norepinephrine (NE) contents of hippocampus and hypothalamus and pain threshold were investigated. The results showed: (1) After ip ACTH, the pain threshold, the contents of 5-HT of the two brain regions and the NE content of hippocampus were markedly elevated. Prior destruction of periaqueductal gray (PAG), the elevation of pain threshold and the increase of the 5-HT contents of two brain regions due to ip ACTH were completely abolished, while the effect of ACTH in elevating NE content of hippocampus still persisted. (2) After intra-PAG injection of ACTH, the pain threshold and the 5-HT contents in hippocampus and hypothalamus were significantly increased, however, the NE levels in hippocampus and hypothalamus showed no significant changes. The analgesic effect of the intra-PAG injection of ACTH was prevented by icv LSD, but not by naloxone, atropine, hexamethonium and phentolamine. (3) After icv ACTH, the pain threshold did not change. These results suggest that the serotoninergic system may be activated by PAG for mediation of ACTH-induced analgesia.