Elevated muscle enzymes and muscle biopsy in idiopathic hypoparathyroidism patients.

OBJECTIVE: To study idiopathic hypoparathyroidism (IHP) myopathy and its pathogenesis by histology method. METHODS: We retrospectively analyzed the clinical data of nine IHP patients during the years 2006-2010. RESULTS: We found that: 1) there was an inverse relationship between the decreased serum calcium level and the elevated muscle enzymes level; 2) the IHP patients had mild-to-moderate muscle cells vacuolar degeneration and focal hyaline degeneration by hematoxylin-eosin (HE) staining; 3) except for 2 patients, the others' muscles striations were undetectable by phosphotungstic acid hematoxylin staining; 4) 2 patients with extremely high level of creatine kinase and relatively longer disease duration had muscle atrophy, multiple focal muscle fiber hyaline degeneration and sarcolemma cells hyperplasia by HE staining; 5) all patients had different degree of immune complex deposition along the muscle cell membranes by immunofluorescence staining. CONCLUSION: The patients with IHP had some histological changes in skeletal muscles but with no specificity. The changes in muscles and the elevated serum muscle enzymes were related to hypocalcemia. The severity of changes in muscles was related to the duration of hypocalcemia and not only to its degree.

Fabrication of ordered NiO coated Si nanowire array films as electrodes for a high performance lithium ion battery.

Highly ordered NiO coated Si nanowire array films are fabricated as electrodes for a high performance lithium ion battery via depositing Ni on electroless-etched Si nanowires and subsequently annealing. The structures and morphologies of as-prepared films are characterized by X-ray diffraction, scanning electron microscopy, and transmission electron microscopy. When the potential window versus lithium was controlled, the coated NiO can be selected to be electrochemically active to store and release Li+ ions, while highly conductive crystalline Si cores function as nothing more than a stable mechanical support and an efficient electrical conducting pathway. The hybrid nanowire array films exhibit superior cyclic stability and reversible capacity compared to that of NiO nanostructured films. Owing to the ease of large-scale fabrication and superior electrochemical performance, these hybrid nanowire array films will be promising anode materials for high performance lithium-ion batteries.

[Construction of T-vectors for the direct, unidirectional cloning and analysis of PCR-amplified promoters].

The amplification and cloning of promoters are regularly employed procedures to study the mechanism of gene regulation. In the present study we developed a method to construct T-vectors used for the direct and unidirectional cloning and analysis of promoters. These so-called T-vectors pEGFP-T and pGL3-T were derived from their parent promoterless vectors pEGFP-1 and pGL3-basic, respectively. To construct the T-vectors, an AhdI recognition site within the Ampr gene in pGL3-basic was silent mutated using overlap extension PCR. Then, a specially designed AhdI cassette was cloned into the respective parent vectors. The procedures of the T-vector construction involved a strategy to minimize the background of nonrecombinant transformants and to eliminate reverse orientation of the PCR products into the T-vectors. The cloning efficiencies of the two T-vectors were both above 85% when tested with a PCR product amplified from a sequence that was pre-confirmed to be able to initiate transcription, and moreover, the constructs harbored the inserts in a desired orientation at a >90% rate. In transient transfection assays, we demonstrated these T-vectors are functional. Thus, the present study provides an easy method to construct a series of T-vectors used for promoter characterization.

Parathyroid hormone regulates osterix and Runx2 mRNA expression predominantly through protein kinase A signaling in osteoblast-like cells.

Runt-related transcription factor 2 (Runx2) and osterix are osteoblast-specific transcription factors essential for the development of osteoblastic cells and bone formation. PTH given intermittently has anabolic effects on bone; however, the exact role remains to be understood completely. The purpose of this study was both to investigate whether PTH regulates Runx2 as well as osterix expression and to identify the signaling used. Using RT-PCR, we confirmed that PTH (1-34) regulated Runx2 and osterix mRNA expression, in rat osteoblast-like cell line UMR 106, in a dose- and time-dependent manner. PTH in low concentrations stimulated both Runx2 and osterix mRNA expression while that in high concentrations did not. Forskolin, an adenylate cyclase activator, also enhanced Runx2 and osterix transcription, and the stimulatory effects of PTH and forskolin were blocked by the pre-treatment of the cells with H-89, a protein kinase A (PKA) inhibitor. In contrast, the protein kinase C (PKC) activator phorbol 12-myristate 13-acetate (PMA) had no effect on Runx2 transcription, but induced an increase in osterix mRNA level at the concentration of 500 nM at 12 h after treatment. Moreover, pre-treatment of the cells with calphostin C, a PKC-specific inhibitor, reduced the increase in osterix transcripts enhanced by PTH and PMA 12 h after treatment. However, these inhibitory effects were not sustained for longer terms. These observations demonstrate that PTH stimulates Runx2 and osterix expression in vitro, at least in part, at transcriptional level. Induction of Runx2 mRNA is mediated through the activation of cAMP/PKA signal transduction. In the case of osterix, although the increase in mRNA level is predominantly mediated via cAMP/PKA signaling, PKC activation might also be involved in this process.

[Studies on the synthesis and bioactivity of alpha-alkylaminobenzyl-phosphonic acids].

AIM: To search for some substituted benzyl phosphonic acids as leading compounds with inhibiting effect on osteoclast formation. METHODS: Target compounds were prepared from aromatic aldehydes, primary amine and phosphorous acid using tetramethylenesulfone as solvent via Arbuzov type reaction. The effect on inhibiting the formation of osteoclast-like cells (OLC) of related compounds was studied by incubating the extract of rat femur marrow. RESULTS: Ten compounds of alpha-alkylaminobenzyl phosphonic acids have been synthesized and identified by MS or 1HNMR analysis. Three (2, 8 and 9) of them were found to have notable effect on the inhibition of OLC formation (P < 0.01). CONCLUSION: Among the present substituted benzyl phosphonic acids, the increased aromaticity and hydrophobicity (such as compound 9) can remarkably enhance the ability to inhibit OLC formation.

Effects of 2-[3-estrone-N-ethyl-piperazine-methyl]tetracycline (XW630) on osteoporosis in ovariectomized rats.

AIM: To study the effects of 2-[3-estrone-N-ethyl-piperazine-methyl]tetracycline (XW630) on experimental osteoporosis in ovariectomized (OVX) rats. METHODS: Serum estradiol (E2) content and bone 1-carboxyglutamic acid-containing protein (BGP) content were measured by radioimmunoassay. With undecalcified bone section and tetracycline intraperitoneal labeling, the bone static and dynamic data were studied in right femur samples. RESULTS: After treatment with XW630 2.5 mg.kg-1, serum BGP content increased by 75.7% but there was no change in serum E2 content and uterus weight compared with OVX rats. Compared with OVX rats, the static data of trabecular bone volume/total tissue volume, trabecular bone volume/sponge bone volume, and mean trabecular plate density were enhanced after treatment with XW630 for 13 wk. The dynamic data of single-labeled surface, double-labeled surface, trabecular osteoid surface, and bone formation rate in tissue level in XW630 group were increased and osteoid maturation period was shortened. CONCLUSION: XW630 enhanced bone activation frequency and increased trabecular connectivity, stability, and strength. XW630 stimulated bone formation and inhibited bone resorption with no effect on reproductive system.

[Effects of pollution with fluoride on bone dynamics of periosteum in iliac of domestic pigs].

Studies on bone dynamic parameters of iliac periosteum and endosteum in domestic pigs with fluorosis caused by coal-burning contamination were conducted. Bone dynamics of 12 pigs with experimental fluorosis and nine controls both labelled with double tetracycline was investigated. Results showed the distance between two labels on iliac periosteum of domestic pigs with fluorosis shortened and, rates of mineralization and bone matrix formation slowed. It suggested fluorosis played significantly inhibitory roles in bone formation on the periosteal membrane of pig iliac.

[The morphological changes in trabecular architecture of bone in severe primary hyperparathyroidism].

By histomorphometry bone biopsy specimens were evaluated from 10 patients with severe primary hyperparathyroidism (PHPT). The data suggest that the mean trabecular width and trabecular bone volume were slightly, but not significantly increased. As indices of bone remodeling activity, both trabecular osteoid surface and trabecular resorption surface were significantly increased indeed, indicating high bone turn-over occurred in PHPT. The dynamic study showed that much more trabecular bone surface was covered either with single or double tetracycline labels in PHPT than in control. The overproduction of osteoid in the patients leads to the accumulation of osteoid on the trabecular surface. Moreover, the mean osteoid width seemed to be thicker in PHPT than in control, suggesting the co-existence of osteomalacia, which might be a possible explanation for eucalcemia usually seen in the majority. Part of chinese PHPT patients.

[Trabecular connectivity in primary hyperparathyroidism].

By bone histomorphometry, 10 cases of primary hyperparathyroidism (PHPT) were used to study the trabecular connectivity, which in a few literatures was mentioned to be reserved somehow. Our data suggest that in comparison with the normal control (10 subjects) the nodes (N.Nd), the number of terminus (N.Tm), the total strut length (TSL), the length between the nodes (Nd.Nd), trabecular separation (Tb.Sp) and trabecular thickness (Tb.Th) were significantly increased while no significance was found in N.Nd/N.Tm, Tm.Tm and TBV. The correlation analyses indicate that there is no correlation between the variables in the normal controls. In contrast, however, TBV and Tb.Th showed negative correlations with N.Tm and Tm.Tm. Besides these, there were positive correlations between N.Nd/N.Tm and TBV or Tb.Th, and a negative correlation between N.Nd/N.Tm and Tb.Sp. The results suggest that to certain degree the trabecular connectivity in PHPT be destructed while the three dimensional architecture be reserved. Our data seem to be different from those reported previously. It may be due to the severity of PHPT in our patients.

Dynamic study on iliac trabeculae of normal Chinese.

We studied 41 iliac bone blocks taken by biopsies from normal Chinese who were divided into several groups by age. On the schedule of 3-11-3-5 days, double tetracycline labellings were administered before bone biopsies. Ten microns undecalcified sections were made with Reichert-Jung microtome and analyzed with Zeizz MOP-Videoplan by light and fluorescent microscopy. The results showed that bone formation had a peak value at adolescence and the mean wall thickness decreased gradually with age. Bone dynamics fluctuated with age because of endocrinological changes in the intrinsic environment. The data we obtained may be used as a reference for clinical investigation of various metabolic bone disorders.

Histomorphometric changes in iliac trabecular parameter with aging.

Iliac bone specimens of 49 normal Chinese subjects were studied. A series of bone histomorphometric parameters were analysed, including mean trabecular width, trabecular bone volume, mean trabecular plate thickness, mean trabecular plate density and mean trabecular plate spacing. The data suggest that there are close correlations of these parameters with aging. The correlations are very significant between mean trabecular width and trabecular bone volume and mean trabecular plate thickness.