Association of use of Chinese herbal medicines and the risk of fracture in patients with osteoporosis: a population-based cohort study.

After utilizing a large population-based claims database and the application of propensity score match approach to reduce the confounding effects, we found that the use of Chinese herbal medicines (CHMs) was related to the lower risk of sequent osteoporotic fracture by 27% among the individuals with osteoporosis. The predominant effect was observed in those receiving CHMs for more than two years. INTRODUCTION: Osteoporosis (OS) is a highly disabling condition that can lead to fragility fracture, thus posing greater burdens of functional limitations for the affected individuals. It is unclear if the use of Chinese herbal medicines (CHMs) could reduce the risk of fracture due to OS. This study aimed to investigate the association of CHMs and the subsequent osteoporotic fracture risk among OS patients. METHODS: This longitudinal cohort study used the Taiwanese National Health Insurance Research Database to identify 250,699 newly diagnosed OS patients aged 20 years or older between 1998 and 2010. We recruited 103,325 CHM users following the onset of OS (CHM users) and randomly selected 103,325 subjects without CHM usage as controls (non-CHM users) by propensity score matching according to the demographic characteristics and comorbidities at enrollment. All enrollees were followed until the end of 2012 to record the incidence of osteoporotic fracture. We applied the Cox proportional hazard regression model to compute the hazard ratio (HR) of the risk of osteoporotic fracture. RESULTS: During the 15-year follow-up period, 7208 CHM users and 11,453 non-CHM users sustained osteoporotic fracture, with an incidence rate of 9.26 and 12.96, respectively, per 1000 person-years. We found that CHM users had a significantly reduced risk of osteoporotic fracture compared to non-CHM users (adjusted HR 0.73; 95% confidence interval [CI] = 0.70-0.75). Those treated with CHMs for longer than 730 days had a lower fracture risk by 54%. Some commonly used CHMs, such as Yan hu suo (Rhizoma Corydalis), Huang Qin (Scutellaria Baicale), Jie Geng (Platycodon grandifloras), Xiang Fu (Cyperus rotundus), Hai Piao Xiao (Cuttlebone Sepium), Jia-Wei-Xiao-Yao-San, Ge-Gen-Tang, Shao-Yao-Gan-Cao-Tang, and Du-Huo-Ji-Sheng-Tang, are related to the lower risk of fracture. CONCLUSIONS: The use of CHMs was associated with lower risk of osteoporotic fracture for OS patients, suggesting that it could be integrated into conventional therapy to prevent subsequent bone fracture.

PacBio assembly of a Plasmodium knowlesi genome sequence with Hi-C correction and manual annotation of the SICAvar gene family.

Plasmodium knowlesi has risen in importance as a zoonotic parasite that has been causing regular episodes of malaria throughout South East Asia. The P. knowlesi genome sequence generated in 2008 highlighted and confirmed many similarities and differences in Plasmodium species, including a global view of several multigene families, such as the large SICAvar multigene family encoding the variant antigens known as the schizont-infected cell agglutination proteins. However, repetitive DNA sequences are the bane of any genome project, and this and other Plasmodium genome projects have not been immune to the gaps, rearrangements and other pitfalls created by these genomic features. Today, long-read PacBio and chromatin conformation technologies are overcoming such obstacles. Here, based on the use of these technologies, we present a highly refined de novo P. knowlesi genome sequence of the Pk1(A+) clone. This sequence and annotation, referred to as the 'MaHPIC Pk genome sequence', includes manual annotation of the SICAvar gene family with 136 full-length members categorized as type I or II. This sequence provides a framework that will permit a better understanding of the SICAvar repertoire, selective pressures acting on this gene family and mechanisms of antigenic variation in this species and other pathogens.

Formation and inhibition of cholesterol oxidation products during marinating of pig feet.

Cholesterol oxidation products (COPs), formed during the heating of cholesterol-rich foods, have been shown to cause cancer and coronary heart disease. The objectives of this study were to develop a GC-MS method for the determination of COPs in pig feet meat, skin, and juice during marinating and to study the formation and inhibition of COPs as affected by the incorporation of soy sauce and sugar. Results showed that an HP-5MS column could provide an adequate separation of cholesterol, 5α-cholestane (internal standard), and seven COPs, including 7α-OH, 7ß-OH, 5,6ß-OH, 5,6α-OH, triol, 25-OH, and 7-keto, within 15 min with a temperature-programming method. Most COPs in pig feet meat were generated at a larger amount than in pig feet skin and marinating juice over a 24 h heating period at about 100 °C. The Maillard browning index rose with increasing heating time, whereas the pH showed a slight change in marinated juice. Both reducing sugar and free amino acid contributed to the formation of Maillard reaction products. The incorporation of soy sauce and crystal sugar into fresh juice was effective in inhibiting COPs formation in pig feet, skin, and juice over a 30 min preheating period.

Formation and inhibition of cholesterol oxidation products in tea-leaf eggs during marinating.

The objectives of this study were to develop a GC-MS method for determination of cholesterol oxidation products (COPs) in tea-leaf eggs and study the formation and inhibition of COPs as affected by heating time and various ingredients in marinated juice. The various COPs in egg and juice samples were extracted by a solvent system of chloroform/methanol (2:1, v/v), followed by purification using a silica cartridge and GC-MS for subsequent separation and quantitation, with high recovery ranges from 85.9 to 98.3% and from 83.1-100.1% being obtained for egg and juice, respectively. 5α-Cholestane was shown to be an appropriate internal standard for quantitation. A total of five COPs, including 7-keto, 5,6 ß-EP, 7α-OH, 7ß-OH, and triol, were formed in tea-leaf eggs during marinating, but not in marinated juice. A peak level of total COPs (2272.2 ng/g) was generated in tea-leaf eggs after 24 h of heating, but reduced to 1068.2 ng/g in 48 h. Both the total phenolic and flavonoid compounds in tea-leaf eggs showed a time-dependent increase during marinating and so did the pH and browning index in tea-leaf eggs and juice. The incorporation of soy sauce or black tea leaf into juice was effective in inhibiting COPs formation in tea-leaf eggs, with the latter being more pronounced than the former. The formation of Maillard reaction products during marinating as well as the presence of total phenolic and total flavonoid in black tea leaf was mainly responsible for COPs reduction in tea-leaf eggs.

Gas chromatography-mass spectrometry determination of conjugated linoleic acids and cholesterol oxides and their stability in a model system.

A gas chromatography-mass spectrometry (GC-MS) method was developed to simultaneously separate cholesterol, eight cholesterol oxidation products (COPs), and two conjugated linoleic acids (9-cis,11-trans-CLA and 10-trans,12-cis-CLA) and to evaluate their stability in a model system during heating. Among four capillary columns tested, an Equity-5 column with low-polar stationary phase provided better resolution within 30 min. A high-performance liquid chromatography method was also developed to determine cholesterol hydroperoxides by using a YMC C30 column with diphenyl-1-pyrenylphosphine as fluorescence reagent. No formation of COPs or degradation of cholesterol and CLAs occurred at 100 degrees C, but the levels of COPs rose drastically at 150 degrees C. The first-order rate of cholesterol degradation declined following a rise in CLA concentration. For 0-, 100-, and 500-microg/ml CLA levels, the formation profiles of 7-hydroxycholesterol, 7-ketocholesterol, and 5,6-epoxycholesterol at 150 degrees C were fitted as multiple first-order curves, whereas a single first-order model could adequately describe 7-hydroperoxycholesterol and cholestane-3beta,5alpha,6beta-triol formation. A CLA-to-cholesterol mole ratio of 0.49 was required to prevent cholesterol oxidation at 150 degrees C.

Encapsulation of lycopene extract from tomato pulp waste with gelatin and poly(gamma-glutamic acid) as carrier.

Tomato pulp waste, a byproduct obtained during the processing of tomato juice, has been shown to be a rich source of lycopene. The objectives of this study were to use gelatin and poly(gamma-glutamic acid) (gamma-PGA) as coating materials for the encapsulation of lycopene extract from tomato pulp waste. Initially, lycopene was extracted with supercritical carbon dioxide, followed by microencapsulation using an emulsion system consisting of 4.5% gelatin, 10% gamma-PGA, and 4.8% lycopene extract. Analysis of differential scanning calorimetry revealed that the thermal stability of the coating material could be up to 120 degrees C, with a mean particle size of 38.7 microm based on Coulter counter analysis. The total weight of microencapsulated powder was 617 microg with the yield of lycopene being 76.5%, indicating a 23.5% loss during freeze drying. During storage of microencapsulated powder, the concentrations of cis-, trans-, and total lycopene decreased along with increasing time and temperature. A fast release of lycopene in the powder occurred at pH 5.5 and 7.0, while no lycopene was released at pH 2.0 and 3.5.

Improved high performance liquid chromatographic method for determination of carotenoids in the microalga Chlorella pyrenoidosa.

Microalgae have become an important commercial source of carotenoids and microalgae-derived functional foods are consumed by people worldwide. Therefore, an HPLC method was developed to discern the variety and content of carotenoids in the microalga Chlorella pyrenoidosa. The microalga sample was powdered, extracted, saponified and subjected to HPLC analysis. A mobile phase of methanol-acetonitrile-water (84:14:2, v/v/v) (A) and methylene chloride (100%) (B) with the following gradient elution was developed: 100% A and 0% B in the beginning, maintained for 14 min, decreased to 95% A in 25 min, 75% A in 30 min, 74% A in 35 min, 45% A in 50 min and returned to 100% A in 55 min. A total of 32 carotenoids were resolved within 49 min by using a C30 column with flow rate at 1 mL/min and detection at 450 nm. An internal standard beta-apo-8'-carotenal was used to quantify all the carotenoids. All-trans-lutein was present in exceptionally large amount (125034.4 microg/g), followed by cis isomers of lutein (27975.3 microg/g), all-trans-alpha-carotene (2465.8 microg/g), zeaxanthin (2170.3 microg/g), cis isomers of beta-carotene (2159.3 microg/g), all-trans-beta-carotene (2155.0 microg/g), cis isomers of alpha-carotene (1766.7 microg/g), beta-cryptoxanthin (334.9 microg/g), neoxanthin and its cis isomers (199.7 microg/g), neochrome (65.2 microg/g), auroxanthin (38.5 microg/g) and violaxanthin and its cis isomers (38.1 microg/g).

In vitro effects of mammalian leptin, neuropeptide-Y, beta-endorphin and galanin on transcript levels of thyrotropin beta and common alpha subunit mRNAs in the pituitary of bighead carp (aristichthys nobilis).

Thyrotropin (thyroid stimulating hormone, TSH) is a member of the pituitary glycoprotein hormones, consisting of two dissimilar subunits, alpha and beta. The two subunits are produced by different genes and are regulated independently. We have previously cloned a TSHbeta cDNA from bighead carp pituitary and investigated its gene regulation. We report here the direct effects of mammalian TSH-releasing hormone (TRH), leptin, neuropeptide-Y (NPY), beta-endorphin and galanin on mRNA levels of both TSHbeta and alpha-subunits in the pituitary of bighead carp in vitro. The dispersed pituitary cells of bighead carp were incubated at 25 degrees C for 6 h with different doses of these factors. The relative mRNA levels of TSHbeta and alpha-subunits were estimated by traditional polymerase chain reaction (PCR) analysis and fluorescence real-time PCR analysis. The results revealed that mammalian TRH, leptin and beta-endorphin produced dose-dependent stimulatory effects on mRNA levels of both TSHbeta and alpha-subunits while thyroxine (T4) and mammalian galanin suppressed mRNA levels of both TSHbeta and alpha-subunits. NPY suppressed TSHbeta mRNA level, but stimulated alpha-subunit mRNA level. This study has demonstrated that mammalian TRH, leptin, NPY, beta-endorphin and galanin were active in modulating the steady-state mRNA levels of TSHbeta and alpha-subunits of bighead carp pituitary in vitro. The results suggest that endogenous TRH, leptin, NPY, beta-endorphin and galanin may modulate transcript levels of TSHbeta and alpha-subunits in pituitary of bighead carp.

Molecular cloning of the cDNAs for pituitary glycoprotein hormone alpha subunits of two species of duck and their gene regulation.

The cDNAs encoding pituitary glycoprotein hormone alpha subunits (PGHalphas) of two species of duck (Muscovy duck, Cairina moschata and Pekin duck, Anas platyrhynchos domesticus) were cloned and sequenced to better understand the phylogenic diversity and evolution of PGHalpha molecules in vertebrates. Oligonucleotide primers were designed and used for reverse transcription PCR (RT-PCR) amplification of PGHalpha cDNA fragments from total cellular RNA of pituitary glands. The remaining sequences were completed by rapid amplification of the cDNA ends. The nucleotide sequence of isolated PGHalpha cDNA of both ducks are identical, including 81 bp of 5' untranslated region (UTR), 360 bp of coding region, and 272 bp of 3'-UTR followed by a 13 bp poly(A)(+) tract. The total number of amino acids deduced from the cDNA of the duck PGHalpha is 120 with a signal peptide of 24 amino acids and a mature protein of 96 amino acids. PGHalphas of the ducks (order Anseriformes) share 96% homology of amino acid sequence in signal peptide, and 100% homology in mature proteins with chicken, quail and turkey (order Galliformes). Our data thus demonstrate identical inter-order homology of PGHalpha mature protein in birds. Ten cysteine residues, presumably forming five disulfide bonds within the alpha subunit, and four proline residues, presumably responsible for folding of the molecule, are conserved in the alpha subunit of ducks. Northern blot analysis revealed that PGHalpha mRNA is expressed only in the pituitary. In order to study factors regulating the gene expression of PGHalpha mRNA, duck pituitary fragments were incubated with GnRH, TRH, testosterone, or triiodothyronine (T(3)). GnRH and TRH increased, while testosterone and T(3) decreased, PGHalpha mRNA levels. This is the first report in birds of TRH up-regulation and down-regulation by testosterone and T(3) under in vitro conditions. The present study demonstrates both ducks have the same cDNA nucleotide and deduced amino acid sequences in the PGHalpha subunit, exhibiting identical inter-genus homology within the family of Anatidae. The findings from mRNA expression work suggest that hypothalamic GnRH and TRH up-regulate, while testosterone and T(3) down-regulate, PGHalpha gene expression in ducks.

Transpedicular wedge osteotomy for correction of thoracolumbar kyphosis in ankylosing spondylitis: experience with 78 patients.

STUDY DESIGN: This is a retrospective study of surgical correction of thoracolumbar kyphosis caused by ankylosing spondylitis. OBJECTIVE: To report the surgical results of thoracolumbar kyphosis deformity corrected with transpedicular wedge osteotomy performed by a single surgeon at a university hospital. SUMMARY OF BACKGROUND DATA: There has not been a large series in the literature reporting on results of the Thomasen-type closing wedge osteotomy for correction of kyphosis deformity secondary to ankylosing spondylitis, nor has two-level osteotomy of this type in one patient ever been described. METHODS: From 1991 through 1998, 92 transpedicular wedge osteotomies were performed in 78 patients with ankylosing spondylitis for correction of fixed flexion deformity of the thoracolumbar spine. RESULTS: The mean amount of correction for each level of osteotomy was 34.5 degrees (range, 15 degrees -60 degrees ). The largest amount of overall correction for a single patient was 100 degrees. Most of the osteotomies (64 of 92) were done at L2 and L3. Fourteen patients with severe deformity required staged two-level osteotomy. Excellent and good results were obtained in 77 patients (98.7%) at the final follow-up. There was no mortality, nor were there any major neurological complications. CONCLUSIONS: Transpedicular wedge osteotomy can effectively and safely correct kyphotic deformity of the thoracolumbar spine caused by ankylosing spondylitis, regardless of rigidity of the spinal curves. Two-level osteotomy can provide sufficient correction for severe cases.