Despite numerous studies have demonstrated an association of type 2 diabetes mellitus (T2DM) and osteoporosis, the underlying mechanism connecting these two conditions remains elusive. Clinically, combined calcium and vitamin D supplement is the commonest osteoporosis therapy; however, recent studies have suggested an increase in cardiovascular risks associated with calcium plus vitamin D supplementation. Therefore, an alternative strategy in treating osteoporosis patients with T2DM is urgently needed. In this study, we hypothesized that combined administration of menaquinone-4 (vitamin K₂, biologically active form of vitamin K) and 1α,25-dihydroxyvitamin D₃ (vitamin D₃, biologically active form of vitamin D) as a novel therapy in treating osteoporosis of T2DM patients. Anabolic effect of vitamin K₂ and vitamin D₃, alone or in combination, was assessed on primary osteoblasts harvested from the iliac crests of C57BL/KsJ lean (db⁺/m⁺) and obese/diabetic (db⁺/db⁺, leptin receptor-deficient) mice. Furthermore, the underlying cellular mechanism was also investigated. Serum undercarboxylated osteocalcin (an indication of vitamin K₂ level) level was higher whereas vitamin D₃ level was lower in db⁺/db⁺ mice, and sections of the iliac crests of db⁺/db⁺ mice illustrated extensive porous structures filled with enlarged adipocytes compared with db⁺/m⁺ mice. Lower levels of bone anabolic markers and bone formation transcription factors (osteocalcin, Runx2, Dlx5, ATF4, type I collagen, OSX, alkaline phosphatase (ALP) activity, p-Smad1/5/8 and p-ERK1/2) were observed in the osteoblasts of db⁺/db⁺ mice. Acute vitamin D₃ (10 nM) application elicited a more sustained and greater magnitude of increase of [Ca²⁺]ᵢ in osteoblasts of db⁺/m⁺ mice when compared with db⁺/db⁺ mice. A significantly higher level of calcium deposits in osteoblasts was observed in db⁺/m⁺ mice when compared to db⁺/db⁺ mice. Co-administration of vitamin K₂ (10 nM) and vitamin D₃ (10 nM) caused an enhancement of calcium deposits in osteoblasts in both strains of mice. Vitamins K₂ and D₃ co-administration time-dependently (7, 14 and 21 days) increased the levels of bone anabolic markers and transcription factors for bone formation, with a greater magnitude of increase observed in osteoblasts of db⁺/db⁺ mice. Suppressed expression of calcium-sensing receptor (CaSR), F-actin, V-ATPase, vitamin D receptor (VDR) and pregnane X receptor (PXR) observed in osteoblasts of db⁺/db⁺ mice were partially reversed by combined vitamins treatment. Moreover, combined vitamins K₂ plus D₃ treatment significantly enhanced migration and the appearance of surface microvilli and ruffles of osteoblasts of db⁺/db⁺ mice. Effects of combined vitamins K₂ plus D₃ treatment observed in osteoblasts of db⁺/db⁺ and db⁺/m⁺ mice were eradicated by warfarin (20 µM, a vitamin K epoxide reductase inhibitor). Thus, our results illustrate that vitamins K₂ plus D₃ supplementation is a novel therapeutic strategy in treating osteoporosis of T2DM patients. / 儘管大量研究已證明第二類型糖尿病和骨質疏鬆症的關聯，連接這兩個病症的基本機制仍然是難以捉摸的。在臨床上，鈣和維生素D的綜合補充劑是最常見的骨質疏鬆症治療，然而最近的研究卻表明服用鈣和維生素D的綜合補充劑會增加患者的心血管風險，因此急切需要尋找可以給予同時患有骨質疏鬆症和第二類型糖尿病患者的替代治療。在本研究中，我們假設甲萘醌-4（維生素K₂，維生素K生物活性形式）和1α，25 - 二羥基維生素D₃（維生素D₃，維生素D的生物活性形式）可以嘗試在同時患有骨質疏鬆症和第二類型糖尿病患者身上作為一種革新的療法。本研究從C57BL/KsJ瘦削/非糖尿病 (db⁺/m⁺) 的小鼠和肥胖/帶有第二類型糖尿病基因 (db⁺/db⁺) 兼有瘦素受體缺陷的小鼠的髂嵴原始成骨細胞上對維生素K₂和維生素D₃單獨或組合使用的合成代謝作用進行了評估。此外，我們也對該成骨細胞的底層機制進行了一系列的研究。 / 在肥胖/帶有第二類型糖尿病基因的小鼠血清內低羧骨鈣素水平（維生素K₂水平的指標）較高而維生素D水平較低，另外，它們的髂嵴的部分與瘦削/非糖尿病的小鼠相比，呈現出比較廣泛的多孔結構並填滿了擴大的脂肪細胞。從肥胖/帶有第二類型糖尿病基因的小鼠的成骨細胞中，可以觀察到它們的骨合成代謝的標誌物和骨骼形成的轉錄因子 (骨鈣蛋白，Runx2，Dlx5，ATF4，第一類型骨膠原，OSX，鹼性磷酸酶 (ALP) 活性，p-Smad1/5/8和p-ERK1/2) 的水平比較低。急性維生素D₃ (10 nM) 的應用在瘦削/非糖尿病小鼠的成骨細胞比起在肥胖/帶有第二類型糖尿病基因的小鼠的成骨細胞引起更持續和更大幅度的細胞內鈣變化增加。在瘦削/非糖尿病小鼠的成骨細胞中比起在肥胖/帶有第二類型糖尿病基因的小鼠的成骨細胞有顯著較高的鈣沉積形成。維生素K₂ (10 nM) 和維生素D₃ (10 nM) 的綜合藥在兩種小鼠的成骨細胞中可以有效地增強鈣沉積的形成。維生素K₂和維生素D₃的綜合藥對增加骨合成代謝的標誌物和骨形成轉錄因子的水平有時間依賴性 (7，14和21日)，療程越長至21日，在肥胖/帶有第二類型糖尿病基因小鼠的成骨細胞中有更大的幅度的增加。合併維生素治療能部分有效地逆轉在肥胖/帶有第二類型糖尿病基因小鼠的成骨細胞中被抑制表達的鈣敏感受體 (CASR)，F-肌動蛋白，V-ATP酶，維生素D受體 (VDR) 和孕烷X受體 (PXR)。此外，結合維生素K₂加維生素D₃治療顯著增強了肥胖/帶有第二類型糖尿病基因小鼠的成骨細胞的細胞遷移和增加了成骨細胞表面外觀的微絨毛和褶皺。在瘦削/非糖尿病小鼠的成骨細胞及肥胖/帶有第二類型糖尿病基因的小鼠的成骨細胞上結合維生素K₂加維生素D₃的治療效果被華法林 (20 μM，維生素K環氧化物還原酶抑製劑) 根除。因此，我們的結果証明了維生素K₂加維生素D₃補充劑的結合使用可有效地作為治療第二類型糖尿病患者並患有骨質疏鬆症的一種新的治療策略。 / Poon, Chui Wa Christina. / Thesis (Ph.D.)--Chinese University of Hong Kong, 2014.n5203 / Includes bibliographical references (leaves 135-151). / Abstracts also in Chinese. / Title from PDF title page (viewed on 26, October, 2016). / Detailed summary in vernacular field only. / Detailed summary in vernacular field only.

Not available / text

Considerable interest exists with regard to the putative therapeutic role of ascorbic acid in various conditions. A condition which has received much attention is cancer, as it is reported that ascorbic acid may be a prophylactic against cancer development. However, the actual involvement of ascorbic acid, an oxidizing/reducing agent, in the development and progression of tumours is presently a subject of much speculation. This study initially addressed the effect of ascorbic acid supplementation over a nutritional concentration range (0 - 100 μg/ml) on the in vitro growth of non-malignant LLCMK and malignant B16 cells. Ascorbic acid supplementation of these two cell types resulted in an overall decrease in the growth of both types of cells. The actual inhibitory mechanism of ascorbic acid on cell growth was not clear. Further study attempted to define and explain a mechanism responsible for this effect. Ascorbic acid has a role in the maintenance of tissue integrity and host defences, thus providing a rational basis for examining its relationship to cancer. Ascorbic acid is lcnown to be essential for the structural integrity of the intercellular matrix of the cells, the latter being a complex aqueous gel containing, amongst other compounds, fats and prostaglandins. Fats and prostaglandins have diverse effects on. membrane stability, enzyme activity and secondary messengers within cells. Hence, this study investigated the effect of ascorbic acid supplementation on certain enzymes and secondary metabolites within the cells, which had the potential to be involved in the control of cell growth. Throughout this study, emphasis was placed on the Bl6 melanoma cells as ascorbic acid supplementation did not significantly affect levels of secondary metabolites within the non-malignant LLCMK cells. Ascorbic acid supplementation of the B16 cells resulted in significant increases in adenylate cyclase activity and cyclic adenosine monophosphate levels, witb a significant decrease in Bl6 cell growth in that particular experiment. As cyclic adenosine monophosphate has a regulatory role in the cell cycle, this study suggested that the inhibitory effect of ascorbic acid supplementation on cell growth was mediated tbrough a final effect provided by the second messenger, cyclic adenosine monophosphate. However, clarification of tbe mechanism of tbe effect of ascorbic acid on adenylate cyclase activity was required. Hence, a further study investigated prostaglandin E₂ levels, as tbese affect adenylate cyclase activity. Prostaglandin E₂ levels were also found to be inversely related to Bl6 cell growth with ascorbic acid supplementation. It thus appeared tbat adenylate cyclase activity was dependent on prostaglandin E₂ levels in the B16 cells, and further study showed that tbis was indeed the case. Here, higher levels of prostaglandin E₂ supplementation of the Bl6 cells inhibited cell growth significantly and also significantly increased adenylate cyclase activity. Arachidonic acid is the precursor of prostaglandin E₂. In the presence of ascorbic acid supplementation, the percentage arachidonic acid composition of the Bl6 cells was inversely correlated with cell growth. Hence, prostaglandin E₂ levels in ascorbic acid supplemented B16 cells appeared dependent on tbe amount of precursor present. This was confirmed when Bl6 cells were supplemented with arachidonic acid. The latter had an inhibitory effect on Bl6 cell growth and also stimulated prostaglandin E₂ production. The cause of tbe inverse relationship between B16 cell growth and arachidonic acid composition with ascorbic acid supplementation was furtber investigated and found to be dependent on tbe uptake of arachidonic acid and other essential fatty acids from tbe medium. The enzymes phospholipase A₂ delta-5 and delta-6-desaturase, and elongase which could influence arachidonic acid levels were not affected to any extent by ascorbic acid supplementation and therefore did not influence the inverse relationship between B16 cell growth and arachidonic acid. Hence, it can be concluded that the effect of ascorbic acid supplementation on the BI6 cells is mediated, in part at least, by cyclic adenosine monophosphate. However, this is not the result of a direct effect of ascorbic acid supplementation. The initial effect of ascorbic acid supplementation concerns fatty acid - in particular arachidonic acid - uptake from the medium, with subsequent cascade effects On secondary metabolites, ultimately affecting the cellular levels of cyclic adenosine monophosphate.

Patients with end-stage renal disease undergoing hemodialysis have a high incidence of oxidative stress-related diseases. This study evaluated oxidative stress and inflammatory markers in patients undergoing hemodialysis before and during vitamin E supplementation. Blood samples were obtained before and after dialysis during two separate dialysis sessions to establish baseline measurements. For the next two months, subjects consumed 400 IU RRR-α-tocopherol daily. At one month and two months of supplementation, blood samples were also obtained before and after dialysis. Circulating concentrations of α- and γ-tocopherols and their metabolites (carboxyethyl-hydroxychromans, α- and γ-CEHCs), vitamin C, and uric acid were determined by HPLC with electrochemical detection. C-reactive protein (CRP), tumor necrosis factor-α (TNF-α), and interleukin-6 (IL-6) were measured using standard clinical assays. F₂-isoprostanes were evaluated using an enzyme immunoassay. Dietary vitamins E and C were assessed using two 24-hour recalls. In response to vitamin E supplementation, plasma α-tocopherol concentrations increased from 18 ± 1.7 μM to 31 ± 5.4 μM (p<0.0001), while γ-tocopherol concentrations decreased from 2.8 ±1.0 μM to 1.7 ± 0.6 μM (p=0.001). Additionally, serum vitamin E metabolites increased, α-CEHCs from 68 ± 20 pmol/ml to 771 ± 161 (p<0.0001) and γ-CEHC from 837 ± 161.8 pmol/ml to 1136 ± 225.9 (p=0.0083). Both CEHCs are well above reported normal values (p<0.0001). Dietary antioxidants (vitamins E and C) were low in most subjects; thus, plasma ascorbic acid levels were low in most subjects, but high in a few, resulting a wide range of responses (88 ± 84 μM). Nonetheless, ascorbic acid concentrations decreased significantly after dialysis to 33 ± 34 μM (p=0.0124), but were unaffected by vitamin E supplementation. Indeed, many parameters decreased significantly by dialysis but were unchanged by vitamin E supplementation, including plasma concentrations of uric acid and TNF-α. Both IL-6 and F₂-isoprostane concentrations were elevated in the subjects but were unaffected by either vitamin E supplementation or dialysis. CRP increased significantly after dialysis (p=0.0161, ANOVA main effect), but in the vitamin E supplemented subjects CRP concentrations were slightly lower before dialysis , but increased following dialysis (p=0.0041, ANOVA interaction). Taken together, the data suggest that there is a complex relationship between chronic inflammation and oxidative stress. Longer supplementation with vitamin E might be necessary in order to observe beneficial effects. / Graduation date: 2003

The present study investigated whether vitamin E supplementation reduced oxidative stress in erythrocytes and improved vitamin E status in patients undergoing hemodialysis (HD). Plasma and erythrocyte α-tocopherol, plasma ascorbic acid, and iron status were determined in 11 regular HD patients prior to and post-dialysis, before and during oral supplementation of vitamin E, 400 IU daily for two months. HD patients were categorized into two groups according to their plasma ascorbic acid levels. We found that only the vitamin C sufficient group (>40 μM, Group I) had reliable measurements of erythrocyte α-tocopherol concentrations before vitamin E supplementation. In Group I prior to dialysis, erythrocyte α-tocopherol concentrations increased in response to vitamin E supplementation from 6.7 ± 0.7 μmol/L packed cells to 9.8 ± 0.6 (μmol/L packed cells (p<0.04). Moreover, there was a positive correlation (p<O.001) between plasma and erythrocyte α-tocopherol levels in Group I subjects. Additionally, vitamin E supplementation significantly increased hematocrits (39.9% ± 1.9% to 42.3% ±1.6%, p<0.004) post-dialysis only in Group I subjects. On the contrary, there was no change in hematocrits during vitamin E supplementation in the vitamin C deficient group (<40 μM, Group II). With respect to measures of iron status and recombinant human erythropoietin (rHuEPO) dose, no differences between before and during vitamin E supplementation were observed in two groups ofpatients. In summary, our data suggest that oral vitamin E supplementation protected erythrocytes from oxidative stress and improved vitamin E status in HD patients, but only in patients with adequate vitamin C status. / Graduation date: 2003

Vitamin A is known to have an important role in cellular differentiation and proliferation. In addition to regulating normal cellular processes vitamin A has also been shown to possess potent antineoplastic activity. The work in this dissertation characterizes the role of retinoic acid (RA) in cellular transformation and tumorigenesis with known oncogene involvement. These studies were initiated by examining the effects of RA on human carcinoma cell lines which express an activated c-ras gene. The bladder carcinoma, EJ/T24 (c-rasᴴ) and the two lung carcinoma cell lines, LXl (c-rasᴷ) and A2182 (c-rasᴷ), were not sensitive to RA. No inhibition of anchorage- or density-dependent growth was observed. Therefore, since these in vitro markers of transformation indicated a lack of effectiveness of RA on carcinomas containing a c-ras gene, retrovirally transformed cells were tested for RA sensitivity. Kirsten murine sarcoma, Balb/c murine sarcoma virus, and Simian sarcoma virus transformed NIH/3T3 and NRK cells were used in these studies. In contrast to the human carcinoma cell lines, anchorage-independent growth of some of the virally transformed cells was very sensitive to inhibition by RA. Anchorage-independent growth of KNRK and SSVNRK cells was sensitive to high concentrations (5 μM) of RA; whereas, Balb/cMSV3T3 and SSV3T3 were sensitive to 1-20 nM RA. BALB/cMSVNRK anchorage-independent growth was stimulated 3.5 fold by 1 μM RA. KNRK displayed a 60% reduction in anchorage-dependent growth at 10 μM RA while little inhibition was observed with the other retrovirally transformed cells. A high level of sensitivity to RA inhibition of anchorage-independent growth was correlated with the presence of cytoplasmic retinoic acid binding protein (CRABP). This indicated that CRABP may have some role in the inhibition of retrovirally induced cellular transformation. RA was shown to significantly reduce the incidence and size of Balb/cMSV3T3 cell tumors in nude mice. The inhibition of tumorigenesis in vivo therefore confirmed the results observed in vitro. To investigate the mechanism by which RA was acting to inhibit retroviral transformation, v-onc mRNA levels were examined. RA had no effect on v-onc mRNA levels in cell lines sensitive to the inhibition of transformation. The effect of RA on the relative rate of synthesis of p21, the transforming protein of KMSV and Balb/cMSV, was investigated. No effect of RA was observed in any of the cell lines. Also, GDP binding by p21 in KNRK cell was unchanged by RA treatment indicating that the functional activity of this transforming protein was not modified. RA does appear to be effective in inhibiting retrovirally induced cellular transformation and tumorigenesis. Evidence presented here indicates that this inhibition is not due to a direct effect of RA on the expression of the v-onc gene and/or gene product. Therefore, some other essential cooperating event(s) occurring within the cell are being acted upon by RA.

Erythrocytic inclusion body syndrome (EIBS) was artificially established in selected stocks of juvenile fall and spring chinook salmon (Oncorhynchus tshawytscha), chum salmon (0. keta), coho salmon (0. kisutch), Atlantic salmon (Salmo salar), and rainbow trout (0. mykiss). Adult spring chinook salmon were also artificially infected with the EIBS virus. Adult male chinook had higher prevalences of EIBS inclusion bodies than females. Cytoplasmic inclusion bodies that are associated with EIBS were not observed in steelhead (0. mykiss), brown (Salmo trutta) nor brook (Salvelinus fontinalis) trout suggesting that these stocks are less susceptible to the EIBS virus. Coho salmon with EIBS were more susceptible to Flexibacter psychrophilus, the causative agent of cold water disease (CWD) than fish without EIBS. The fish with EIBS were most susceptible to F. psychrophilus during the first 20 days after virus exposure, when inclusion bodies were most prevalent. Coho salmon infected with both the EIBS virus and F. psychrophilus required a longer recovery period than fish exposed to either pathogen alone. Most investigations of EIBS require in vivo experimentation and artificial infections using diseased fish tissues. Heterologous tissue used to establish EIBS did not contribute to anemia nor mortality. Death was not attributed to the EIBS virus alone but to the combined effects of the virus and a secondary pathogen. The severity of EIBS may be reduced with dietary Vitamin C prophylaxis. Fish fed 1,000 mg ascorbic acid/ Kg of diet had the fewest signs of EIBS; they had the highest hematocrit values and the lowest incidence of cytoplasmic inclusion bodies. However, vitamin C therapy alone was not sufficient to prevent the disease. / Graduation date: 1992

Objective: Acute lower respiratory infection (ALRI) is the leading cause of mortality in pediatric group all around the world. Vitamin D has been demonstrated to play a possible role in the prevention of ALRI in children because of its physiological importance in the immune system. This systematic review aims to explore the protective role of vitamin D on ALRIs among children and its preventive effectiveness by synthesizing RCTs. And the other objective is to determine dosage of vitamin D with the best effect by investigating the association of different level of vitamin D supplementation with risk of ALRIs. Methods: Studies were searched through three databases PubMed, ISI Web of Knowledge and Cochran Central Register of Controlled Trials and Cochran Library databases among publication from April2003 to April 2013 with a combination of key terms. Inclusion and exclusion criteria were used to select studies. And then CONSORT guideline and JADAD scale were used to assess quality of these studies. Data on outcome measurements including health outcomes (e.g. incidence of pneumonia and influenza A, duration of recovery of pneumonia and bronchiolitis, the risk of relapse of pneumonia, the number of parent-reported ARIs); and surrogate outcomes (e.g. measuring scores of ATAQ test) were extracted and tabulated. The association with vitamin D level of risk of ALRIs were explored as well. Results: Eight RCTs were found to be relevant and adopted in this systematic review of the 796 identified articles in English or Chinese. The findings were mixed, but most studies suggested vitamin D supplementation reduced risk or illness duration of ALRIs significantly among children with different levels of vitamin D deficiency. Four studies suggested statistically significant risk reduction on incidence of repeat pneumonia (by29%, 95%CI 6% to 46%), parent-reported ARIs (by 48%, 95%CI 11% to 69), influenza A (by 42%, 95%CI 1% to 66%), and asthma exacerbation triggered by ALRIs (P= 0.029), while one study showed an insignificant outcome. For recovery events and hospitalization of ALRIs, three studies suggested statistically significant reduction on recovery time from pneumonia (P= 0.008), severe asthma (P= 0.004) and bronchiolitis (P< 0.05), and two studies suggested significant decrease on duration of hospitalization for bronchiolitis (P< 0.05) and pneumonia (P= 0.005). The increasing changes in serum 25(OH)D were consistent with the significant difference of ALRIs events between intervention and control groups. Conclusion: Overall, the evidence is insufficient to conclude that vitamin D supplementation is beneficial to all kinds of children in preventing or assistant treating ALRIs. More number of high quality, large scale and unbiased RCTs should be conducted to confirm the effectiveness of vitamin D among children in Hong Kong and different areas in mainland China. / published_or_final_version / Public Health / Master / Master of Public Health

Not available / text

Not available / text