BACKGROUND: The pathological changes of osteoarthritis are similar to that of kaschin-beck disease, but the cause of the latter is still unclear,damage is more serious. From the comparison of pathology changes between them, we hope to find out an effective therapeutic method with the assistance of arthroscope basing on the experiences in osteoarthritis.OBJECTIVE: To make comparison of the histomorphological changes between osteoarthritis and kaschin-beck disease with knee arthroscope,which give clues to the diagnosis and selection of operative technique for kaschin-bock disease under arthroscope.DESIGN: Case control study.SETTING: Department of Orthopaedics, Shaanxi People's Hospital.PARTICIPANTS: Thirty-five patients who needed arthroscope surgery due to severer knee osteoarthritis were randomly selected from Department of Orthopaedics, Shaanxi People's Hospital between April 1998 and September 2000; Meanwhile 31 patients with knee joint functional obstacle caused by kaschin-beck disease (epidemic disease area) were randomly selected from Luochuan Hospital of Shaanxi Province in November 2001.METHODS: Japanese made OLYMPUS series color television arthroscope and surgery system were used to carry out knee joint operations in all participants. During the operation, video recording, postoperative editing, photographs, comparison and analysis were conducted, in order to find out the pathological changes of the two groups in knee joint synovium, patella, femur, tibia and meniscus with the assistance of arthroscope.RESULTS: Totally 66 patients were enrolled in this study and all endamage in Kaschin-beck disease and osteoarthritis: Injuries were graded into grade 0, 1, 2 and 3, there were 0,3,5,23 cases and 0,3,16,16cases correspondingly in kaschin-beck disease group and osteoarthritis cartilage damage was presented by big piece of cartilage peeled off, part of them came off leading to bone exposure, the exposed bone surface is uneven; while cartilage damage in osteoarthritis was presented by the wear of cartilage, moreover cartilage island was found remained in the exposed bone with the surface covered with fiber strips and the exposed observed in 24 patients with Kaschin-beck disease, 5 cases with coarse stress point and 19 cases with burr-like damage, amongst which there were 9 cases displayed both medial and lateral meniscus damages; in osteoarthritis group there were 16 cases showed meniscus damages including edge wears in11 cases and breakage in 5 cases.CONCLUSION: Cartilage damage in knee articular was obviously severer in Kaschin-beck disease than in osteoarthritis, although kaschin-beck disease is no matter resulting in serious knee joint damage, but its pathology changes are different from osteoarthritis, we should pay more attention to the peculiarity of big piece of cartilage peeling off during operations.

There is a closely relationship between vitamin K and osteoporosis. As a cofactor for carboxylase activity, vitamin K can facilitate the conversion of glutamyl to gamma-carboxyglutamyl residues and influence the synthesis and excretion of gamma-carboxylation of osteocalcin to increase the formation of bone. Vitamin K can also effectively inhibit the absorption of bone mass. Besides, there are increasing evidences that vitamin K can effect the synthesis and excretion of nephrocalcin and interlukin-1,6 that can regulate calcium balance and bone metabolism. Meanwhile, there is a consistent line of evidence in human epidemiologic and intervention studies that clearly demonstrate that vitamin K can not only increase bone mineral density in osteoporotic people, but also reduce fracture rates to improve bony health. However more researches are required before vitamin K is widely applied in prevention and treatment of osteoporosis. The American Medical Association recently has increased the dietary reference intakes of vitamin K to 90 mg/d for females and 120 mg/d for males.
Animals ; Bone Density ; drug effects ; Humans ; Nutritional Requirements ; Osteocalcin ; metabolism ; Osteoporosis ; drug therapy ; etiology ; prevention & control ; Vitamin K ; antagonists & inhibitors ; pharmacology ; therapeutic use

In this study, nattokinase gene was amplified by PCR using bacillus subtilis chromosomal DNA as template and cloned into expressed vector pBV220. After transforming recombinant plasmid into E.coli HB101, the recombinant strain was yielded. It was proved that expression products was secretive and expression protein was 12% of total cell protein by SDS-PAGE. Optimum culture time and inducing time was determined as 6h and 5h respectively. The plasmid stability studies showed that recombinant plasmid has excellent segregational stability but the structural stability was not good in the host cell.

Objective:To analyze the activities of human NKG2D ligand MICA/B promoter induced by 5-Fu.Methods:The 5'-end flanking regions of MICA /B promoter and their different truncated fragments were amplified from A549 genome by PCR. The resulting amplicons were cloned into pGL3-Basic vector to generate the MICA/B luciferase reporter plasmids. All the constructs were transiently transfected A549 cells. The promoter region activities were determined by dual-luciferase reporter assays. The effect of 5-Fu on the promoter activities of MICA/B was also tested.Results and Conclusion:The 5'-end flanking regions of MICA /B promoter and five of their different truncated fragments were successfully obtained. The normalized luciferase reporter gene activities driven by the above promoters and fragments were 3.61,2.26,1.63,0.313,0.711 and 0.663 for MICA and 17.49,10.11,7.398,0.822,0.997 and 0.49 for MICB,respectively. Promoter activities in transiently transfected A549 cells treated by 20,40,80,160 and 320 μg/m of 5-Fu increased 1.69,1.48,1.62,1.55 and 1.78 fold for MICA and 1.44,1.87,1.38,1.19 and 1.25 fold for MICB. Our results suggest that 5-FU can significantly up-regulate the promoter activity of both MICA and MICB.

Abstract: Objective　To investigate a poisoning incident caused by eating eight treasure congee, and establish liquid chromatography (LC)-mass spectrometry (MS)/MS screening method of 28 alkaloids to provide references for disposal of similar poisoning incidents. Methods　LC-MS/MS was used for screening 28 alkaloids in the urine, eight treasure congee and food raw material, and the detected alkaloids were quantified. Samples were extracted with 0.4% formic acid aqueous solution and separated by a Acquity UPLC BEH C18 column (1.7 μm, 100 × 2.1 mm). Acetonitrile-0.2% formic acid aqueous solution was used as the mobile phase and gradient elution was adopted. The ionization mode was electrospray positive ionization mode, and the detection method was multi-reaction monitoring (MRM). Analytes were quantified with the external standard method. Results　In the concentration range of 0-100 ng/mL, the linear correlation coefficient r were greater than 0.999 for 28 alkaloids. The recovery of 28 alkaloids in urine sample ranged from 63.0% to 105.0%, and the relative standard deviations (RSDs) were between 5.8% and 8.6%. The recovery of 28 alkaloids in eight treasure congee sample ranged from 72.0% to 109.0%, and the RSDs were between 6.3% and 9.7%. The recovery of 28 alkaloids in semen sesami nigrum sample ranged from 60.0% to 95.0%, and the RSDs were between 4.8% and 8.2%. Hyoscyamine (2 380.0 ng/mL), scopliamine (3.6 ng/mL) and rac-anisodamine (4.7 ng/mL) were detected in the patient's urine. Hyoscyamine (63.3 μg/g), scopliamine (5.7 μg/g) and rac-anisodamine (2.1 μg/g) were detected in eight treasure congee. Hyoscyamine (901.0 μg/g), scopliamine (80.0 μg/g) and rac-anisodamine (30.1 μg/g) were detected in the seed of Datura stramonium L. The ratio of scopliamine and hyoscyamine in the seed of D. stramonium was 1∶11, which complies with the characteristics of D. stramonium L. In urine sample, the proportion of scopliamine and rac-anisodamine was 0.15% and 0.20%, and hyoscyamine accounted for 99.65%. Conclusion　Seed morphology, the content range and proportion of three alkaloids are all in accord with the characteristics of D. stramonium. Combined with the clinical symptoms of atropine poisoning, it can be deduced that this incident is a family food poisoning caused by accidental consumption of seed of D. stramonium L. The method can provide technical support for the clinical diagnosis and treatment of alkaloid poisoning patients, and also provide a basis for emergency detection and disposal of alkaloid poisoning events.

OBJECTIVETo study the content of mineral elements and the principal components in Gastrodia elata.

METHODMineral elements were determined by ICP and the data was analyzed by SPSS.

RESULTK element has the highest content-and the average content was 15.31 g x kg(-1). The average content of N element was 8.99 g x kg(-1), followed by K element. The coefficient of variation of K and N was small, but the Mn was the biggest with 51.39%. The highly significant positive correlation was found among N, P and K . Three principal components were selected by principal components analysis to evaluate the quality of G. elata. P, B, N, K, Cu, Mn, Fe and Mg were the characteristic elements of G. elata.

CONCLUSIONThe content of K and N elements was higher and relatively stable. The variation of Mn content was biggest. The quality of G. elata in Guizhou and Yunnan was better from the perspective of mineral elements.

Drugs, Chinese Herbal ; analysis ; Gastrodia ; chemistry ; Minerals ; analysis ; Principal Component Analysis

This study was designed to use iTRAQ technology coupled with 2D LC-MS/MS to study the comparative proteomics of different processing technology for pilose antler. 1015 proteins were identified with 2D LC combined with MOLDI TOF/TOF mass spectrometry. Comparative analysis with Protein Pilot (Version 4.5) revealed that 87 proteins were changed (P ≤ 0.05, the ratio of > 1.50 or < 0.60 as the threshold selection of difference proteins), of which 24 were up regulated and 33 were down regulated in the traditional frying process (TFP) compared with the fresh pilose antler (P ≤ 0.05). 7 significant different proteins (P ≤ 0.001), most of these significantly changed proteins were found to be involved in calcium ion binding and ATP binding associated with human healthy. Freeze drying with protective agent (FDP) (Trehalose) can improve the content of significantly different proteins (P ≤ 0.001) including Collagen alpha-1 (XII) chain (COL12A1) and Collagen alpha-1 (II) chain (COL2A1). The significant function involves in platelets activating, maintenance of spermatogonium, and disorder expression in tumor cells. The functional annotation by Hierarchical clustering and GO (gene ontology) showed that the main molecule functions of the proteins significantly changed in these processes were involved in binding (52.7%), catalytic (25.3%), structural molecule and transporter (6.6%).
Animals ; Antlers ; chemistry ; Chromatography, Liquid ; Collagen ; chemistry ; Down-Regulation ; Freeze Drying ; Gene Expression Regulation ; Proteomics ; Tandem Mass Spectrometry ; Technology, Pharmaceutical ; methods ; Up-Regulation

OBJECTIVETo identify the expression characteristics of the 1700001022RIK (RIKEN cDNA 1700001022) gene in mice and explore its function by bioinformatic analysis.

METHODSUsing the expression profile of gene microarray, we detected the expression of a new testis-specific gene, 1700001022RIK, in mice. We analyzed its expression characteristics in the testis tissue and their changes in different developmental stages of the testis by RT-PCR, real-time RT-PCR, Western blot, and immunohistochemistry. We performed bioinformatic analysis using a bioinformatic software.

RESULTSThe 1700001022RIK gene was specifically expressed in the mouse testis in an age-dependent manner, most highly in the adult mice. The 1700001022RIK protein was mainly expressed in the spermatogonia, spermatocytes, and round spermatids of the adult mice. Bioinformatic analysis showed that the 1700001022RIK protein amino acid sequence had a high similarity in human and mice, which indicated that this gene was highly conserved in mammals.

CONCLUSION1700001022RIK is a testis-specific gene mainly expressed in the spermatogonia, spermatocytes, and round spermatids of seminiferous tubules, which might be involved in the regulation of spermatogenesis.

Age Factors ; Animals ; Blotting, Western ; Computational Biology ; DNA, Complementary ; Gene Expression ; Genomics ; Male ; Mice ; Molecular Chaperones ; genetics ; Seminiferous Tubules ; Spermatids ; Spermatocytes ; Spermatogenesis ; genetics ; Spermatogonia ; Testis

OBJECTIVE To have a systematic pathomechanism view of three chest impediment-syndromes of Qi Deficiency and Blood Stasis syndrome(QDBS),Qi Stagnation and Blood Stasis syn-drome (QSBS), Cold Obstruction and Qi Stagnation syndrome(COQS) and further investigate the changed metabolome and related pathways for screening potential biomarkers in rat plasma. METHODS According to clinical pathogeny, three kinds of syndrome models were established to simulate the disease of chest impediment. Plasma metabonomics based on UPLC-Q-TOF/MS was applied in this research to detected small molecule metabolites for identifyingthe special potential biomarkers of three chest impediment syndromes, respectively. RESULTS Significant metabolic differences were observed between thecontrol group and three syndrome groups. Furthermore, three syndrome groups were distinguished clearly by pattern recognition method.The particular metabolites contributing most to the classification of three chest impediment syndromes were identified. In the QSBS group, the potential biomarkers could include 2-keto-glutaramic acid, L-methionine, L-homocysteic acid, octadecanamide, stearoylglycine,behenic acid,linoleylcarnitine,lysoPC(14:1(9Z)),indoxyl sulfate and cholic acid.In the COQS group, they could be aminoadipic acid, palmitic amide, oleamide, lysoPC(P-16:0), lysoPC(P-18:0), lysoPC(20:2(11Z,14Z)), 9-HETE and tauroursodeoxycholic acid. Moreover, 4-pyridoxic acid, L-palmi-toylcarnitine, lysoPC(20:0), lysoPC (22:5 (4Z,7Z,10Z,13Z,16Z)), 3- hydroxyhexadecanoic acid and arachidonic acid could be the potential biomarkers for the QDBS group. CONCLUSION Three chest impediment syndromes have their own potential biomarkers.Each special metabolite has its owndifferent metabolic pathway.Both metabolismof cysteine and methionine,and metabolism of alanine,aspartate and glutamate are the main pathways in regulation of metabolic disorders in QSBS syndrome. Lysine biosynthesis and degradation,fatty acid metabolism,and glycerophospholipid metabolism are the main pathways in regulation of metabolic disorders in COQS syndrome.Arachidonic acid metabolism, fatty acid metabolism,fatty acid elongation in mitochondria,and vitamin B6 metabolism are the main pathways in regulation of metabolic disorders in QDBS syndrome.These endogenous substances were indicated as the special potential biomarkers for three chest impediment syndromes and worth studying in depth.

PURPOSE: The incidence and mortality of breast cancer is increasing worldwide. There is a constant quest to understand the underlying molecular biology of breast cancer so as to plan better treatment options. The purpose of the current study was to characterize the expression of histone deacetylases-3 (HDAC3), a member of class I HDACs, and assess the clinical significance of HDAC3 in breast cancer. METHODS: Quantitative real-time polymerase chain reaction, immunohistochemistry, and western blot analysis were used to examine messenger RNA and protein expression levels. The relationships between HDAC3 expression and clinicopathological variables were analyzed. MTT assays were used to detect cell proliferation. Glucose-uptake, lactate, adenosine triphosphate, and lactate dehydrogenase assays were employed to detect aerobic glycolysis. Chromatin immunoprecipitation was used to detect microRNA-31 (miR-31) promoter binding. RESULTS: Our data revealed that HDAC3 was upregulated in breast cancer tissue compared with matched para-carcinoma tissues, and high levels of HDAC3 were positively correlated with advanced TNM stage and N stage of cancer. Furthermore, overexpression of HDAC3 promoted breast cancer cell-proliferation and aerobic glycolysis. The functional involvement of HDAC3 was related in part to the repression of miR-31 transcription via decreased histone H3 acetylation at lysine K9 levels of the miR-31 promoter. Survival analysis revealed that the level of HDAC3 was an independent prognostic factor for breast cancer patients. CONCLUSION: Our findings revealed that HDAC3 served as an oncogene that could promote cell proliferation and aerobic glycolysis and was predictive of a poor prognosis in breast cancer. HDAC3 participated in the cell proliferation of breast cancer, which may prove to be a pivotal epigenetic target against this devastating disease.
Acetylation ; Adenosine Triphosphate ; Blotting, Western ; Breast Neoplasms* ; Breast* ; Cell Proliferation* ; Chromatin Immunoprecipitation ; Epigenomics ; Glycolysis* ; Histone Code ; Histones* ; Humans ; Immunohistochemistry ; Incidence ; L-Lactate Dehydrogenase ; Lactic Acid ; Lysine ; Molecular Biology ; Mortality ; Oncogenes ; Prognosis* ; Real-Time Polymerase Chain Reaction ; Repression, Psychology ; RNA, Messenger