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ObjectiveTo investigate the effect of Yiqi Jiedu prescription-containing serum on the proliferation of medullary thymic epithelial cells （mTEC） and regulatory T （Treg） cells in myasthenia gravis （MG） patients with thymus hyperplasia. MethodAccording to serological methods，35 SD rats were adaptively fed for one week and randomized into the low-，medium-， and high-dose Yiqi Jiedu prescription groups，control group， and prednisone group，with seven rats in each group， which were then gavaged with the corresponding drugs for one week for preparing the drug-containing serum. The effect of Yiqi Jiedu prescription-containing serum at different concentrations on the proliferation of mTEC and Treg cells were determined by cell counting kit-8 （CCK-8） assay. Besides， the effect of mTEC and Yiqi Jiedu prescription-containing serum on Treg cell proliferation were observed through co-culture. ResultThymocytes were cultured for a period of time. Their mean positive rate revealed by flow cytometry using mTEC characteristic marker Ulex europaeus agglutinin Ⅰ （UEAI） was 92.54%. Treg cells were sorted by magnetic beads. The purity of Treg cells after repeated magnetic bead sorting was as high as 92%. mTEC and Treg cells showed high positive expression rates，and their cell purity met the requirements of subsequent experiments. When the concentration of Yiqi Jiedu prescription-containing serum was 2.5%-15%，it exhibited an inhibitory effect against mTEC and Treg cells. When the concentration was equal to or greater than 20%，it promoted cell proliferation，which was further enhanced with the extension of action time. The results after 48 h of culture showed that compared with the control group，prednisone and low-dose Yiqi Jiedu prescription had no significant effect on the proliferation of these two kinds of cells，but the medium- and high-dose Yiqi Jiedu prescription remarkably reduced their proliferation inhibition rate （P<0.01）. After co-culture with mTEC， the control group was not significantly different from the prednisone group and the low-dose Yiqi Jiedu prescription-containing serum group in the proliferation of Treg cells，while the medium- and high-dose Yiqi Jiedu prescription-containing serum groups significantly lowered the proliferation inhibition rate （P<0.01）. ConclusionYiqi Jiedu prescription-containing serum affects the proliferation of mTEC and Treg cells in MG patients with thymus hyperplasia. Compared with the solely cultured Treg cells isolated from MG patients，the Treg cells co-cultured with mTEC exhibit enhanced proliferation in MG patients，suggesting that mTEC can regulate the proliferation of Treg cells. This effect becomes more obvious after the intervention with Yiqi Jiedu prescription-containing serum，indicating that intervention effect of Yiqi Jiedu prescription on Treg cells can be produced during its treatment of mTEC， which may be one of the mechanisms of Yiqi Jiedu prescription-containing serum in alleviating MG.
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Aim To investigate the molecular mechanism of the protection of vascular smooth muscle cells by the regulation of endoplasmic reticulum stress and autophagy by Fufang Danshen dripping pills.Methods Fifty patients with atherosclerosis were randomly divided into two groups; one group received normal treatment,while the other group was added Fufang Danshen dripping pills,and the clinical efficacy was observed.Vascular smooth muscle cells were divided into control,ox LDL model,Fufang Danshen dripping pill group,Fufang Danshen dripping pill+endoplasmic reticulum stress inhibitor group,and Fufang Danshen dripping pill+endoplasmic reticulum stress inducer group.Proliferation was detected,and vasodilator function factors and oxidative stress were measured in each group,ER stress marker proteins,autophagy marker proteins and apoptosis related protein expression were detected,and activation of the IRE1-TRAF2-ASK1-JNK signaling pathway was detected.Results Compared with control group,various indicators of cells in ox-LDL model group showed that they were under ER stress,high oxidative stress,high autophagy status,and the IRE1-TRAF2-ASK1-JNK pathway was found to be over activated.However,compared with ox LDL model group,the above indicators in Fufang Danshen dripping pill group and Fufang Danshen dripping pill+endoplasmic reticulum stress inhibitor group were significantly better,the IRE1-TRAF2-ASK1-JNK pathway was over activated,and the endoplasmic reticulum stress inhibitor was more obvious,and Fufang Danshen dripping pill+endoplasmic reticulum stress inducer group significantly reversed the improved effects of Fufang Danshen dripping pills.Conclusions Fufang Danshen dripping pills protect vascular smooth muscle cells by inhibiting excessive activation of the IRE1-TRAF2-ASK1-JNK pathway,decreasing endoplasmic reticulum stress,maintaining proper autophagy,and inhibiting abnormal cell proliferation and apoptosis.
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This study investigated the effect of rhein(RH) on the apoptosis and autophagy of human umbilical vein endothelial cells(HUVECs) induced by hydrogen peroxide(H_2O_2) and its underlying mechanism. The oxidative damage model in HUVECs was established and the cells were divided into different treatment groups. Cell survival rate was detected by MTT assay, apoptosis by Annexin V-FITC/PI double staining and Hoechst 33258 fluorescence staining, autophagy by Ad-mCherry-GFP-LC3 B adenovirus transfection, and protein expression by Western blot. The results showed that RH could protect cells by increasing the cell survival rate in a dose-dependent manner, decreasing the expression of apoptosis-related proteins(Bax and cleaved caspase-3) and the ratio of Bax/Bcl-2, elevating the expression of Bcl-2, up-regulating the expression of microtubule-associated protein 1 light chain 3(LC3)-Ⅱ, and down-regulating the expression of p62. Adenovirus transfection results showed that RH could increase the green and red spots, as well as the yellow spots. However, after the addition of autophagy inhibitor 3-MA, autophagy was reduced and apoptosis was increased. RH could enhance the expression of silent information regulator 2 related enzyme 1(SIRT1). The addition of SIRT1 inhibitor EX-527 reduced the protective effect of RH and cell viability. The addition of 3-MA had no effect on the expression of SIRT1 protein, but the expression of SIRT1 and LC3-Ⅱ proteins decreased and the expression of p62 increased after the addition of EX-527. After RH treatment, the phosphorylation of adenosine monophosphate-activated protein kinase(AMPK) increased, while that of the mechanistic target of rapamycin(mTOR) decreased in a dose-dependent manner. Moreover, this effect could be weakened by the AMPK inhibitor compound C. RH may enhance autophagy through SIRT1/AMPK/mTOR pathway to reduce H_2O_2-induced apoptosis of HUVECs.
Subject(s)
Humans , Anthraquinones , Apoptosis , Autophagy , Human Umbilical Vein Endothelial Cells , Hydrogen Peroxide , Signal TransductionABSTRACT
OBJECTIVE@#To assess the 10-year Atherosclerotic Cardiovascular Disease (ASCVD) risk score among adults in eastern China using the China-PAR equation which formulated primarily for the Chinese population.@*METHODS@#Data from 72,129 individuals from 35-74 years old who received routine physical examinations in eastern China were analyzed in this study. The 10-year risk scores were calculated using the China-PAR equation. The chi-square test and logistic regression were then performed to evaluate the association between the selected risk factors and overall CVD risk.@*RESULTS@#The mean 10-year ASCVD risk scores were 3.82% ± 3.76% in men and 1.30% ± 1.65% in women based on the China-PAR equation. Overall, 20% of men and 3.5% of women were intermediate-risk, and 7.3% of men and 0.3% of women were high-risk. Waist to hip ratio (WHR) [OR = 1.16 (CI 95% = 1.06-1.26)], waist to height ratio (WHtR) [OR = 1.16 (CI 95% = 1.05-1.28)], non-high-density lipoprotein cholesterol (non-HDL-C) [OR = 1.23 (CI 95% = 1.09-1.39)], and total cholesterol (TC)/HDL-C [OR = 1.68 (CI 95% = 1.46-1.94)] were more strongly associated with CVD risk than body-mass index (BMI), waist circumference (WC), and TC alone.@*CONCLUSION@#Male-specific prevention and treatment strategies for ASCVD are needed in eastern China. In addition, WHR, WHtR, non-HDL-C, and TC/HDL-C which not included in the the China-PAR equation were also independently associated with 10-year ASCVD risk score categories.
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Adult , Aged , Female , Humans , Male , Middle Aged , Algorithms , Atherosclerosis , Epidemiology , China , Physical Examination , Risk FactorsABSTRACT
Background Doxycycline is a broad spectrum antibiotic,and it is frequently used in the treatment of ocular surface diseases.Objective The purpose of the present study was to investigate the effect of doxycycline on the inhibition of cell proliferation in bovine corneal myofibroblasts in vitro and assess its contribution to ocular surface repairing mechanism.Methods Six fresh bovine corneas were collected.The corneal stromal layer was isolated by two-step method of 1.0 g/L and 2.0 g/L collegenase-1.Isolated cells were plated at mantaryay culture flask in 10％ FBS of RPMI-1640.Vimentin and alpha-smooth muscle actin (α-SMA) organization were evaluated by immunocytochemistry,and the cells with influoresccence staining for vimentin and α-SMA were identified as the corneal myofibroblasts.Doxycycline at the concentrations of 10,20,40,60,80 mg/L was added to the medium,respectively,in different concentrations of doxycycline groups.Dexamethasone (120 mg/L)was used in the same way in the positive control group,and no drug was used in the negative control group.Cell proliferation was evaluated by MTT and the cell cycle was analyzed by BD FACScan flow cytometer assay 24 hours and 48 hours after addition of any drug.Results The cells grew well and showed the positive response for vimentin and α-SMA.MTT assay showed that the A570values of bovine corneal myofibroblasts were gradually declined with the increase of the concentration of doxycycline and lapse of active time,showing statistically significant difference (Fconcentration =1233.778,P＜0.001 ; Ftime =227.564,P ＜ 0.001).And the difference between the two factors was also statistically significant (Ftime*concentration =51.656,P＜0.001).Flow cytometry cell cycle analysis showed that 24 hours after 10,20,40,60,80 mg/L doxycycline treated,the perentage of of corneal myofibroblast cell in G0-G1 phase was 82.85％,84.36％,85.18％,87.12 ％ and 89.31％,showing significant increase in comparison with 63.89％ of the negative control group (all P＜0.05),and that of 40 mg/L doxycycline group was near the positive control group.Forty-eight hours after 10,20,40,60,80 mg/L doxycycline treated,the perentage of of corneal myofibroblast cell in G0-G1 phase was 82.78％,86.15％,88.23％,89.57％,93.00％,with significant increase in comparison with 70.17％ of the negative control group (all P ＜ 0.01),and that of 40 mg/L doxycycline group was near the positive control group.Conclusions The growth of the bovine corneal myofibroblasts is inhibited by doxycycline in time-and dosedependent manner in the range from 10 mg/L to 80 mg/L,and 40 mg/L of doxycycline has an obviously inhibitory action as 120 mg/L dexamethasone.
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This study is to investigate whether naked plasmid DNA can effectively transfect lung cancer related cells and express human kallistatin, an endogenous protein that inhibits angiogenesis and tumor growth, and to explore the biological activity of the low-level expressed kallistatin to lung cancer in vitro and in vivo. The plasmids were delivered with Lipofectamine 2000 to transfect various lung cancer related cells. Kal expression was determined by ELISA. The biological effects of Kal expression on proliferation, migration and apoptosis rate of the cells were examined. In subcutaneous NCI-H446 xenograft model, pKal was injected directly into tumors, the changes of CD34, Ki-67 and E-cadherin expression were detected with immunohistochemical assay, the tumor apoptosis was analyzed with TUNEL assay. Both the endothelial cell and lung cancer cells could express kallistatin after plasmid transfection. The proliferation and migration of human umbilical vein endothelial cells were inhibited, but the apoptosis rate was not affected. The proliferation rates of all the three tested lung cancer cells, such as NCI-H446, NCI-H460 and A549, were inhibited, and their apoptosis rates were enhanced, but different cells behaved differently. In subcutaneous NCI-H446 xenograft model, intratumor injection of pKal inhibited the growth of lung cancer by reducing angiogenesis and proliferation of tumor cells. In conclusion, this study demonstrated the efficacy of plasmid-mediated expression of kallistatin to lung cancer related cells, thus providing a basis for their clinical application in the treatment of lung cancer.
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This study is to investigate whether naked plasmid DNA can effectively transfect lung cancer related cells and express human kallistatin, an endogenous protein that inhibits angiogenesis and tumor growth, and to explore the biological activity of the low-level expressed kallistatin to lung cancer in vitro and in vivo. The plasmids were delivered with Lipofectamine 2000 to transfect various lung cancer related cells. Kal expression was determined by ELISA. The biological effects of Kal expression on proliferation, migration and apoptosis rate of the cells were examined. In subcutaneous NCI-H446 xenograft model, pKal was injected directly into tumors, the changes of CD34, Ki-67 and E-cadherin expression were detected with immunohistochemical assay, the tumor apoptosis was analyzed with TUNEL assay. Both the endothelial cell and lung cancer cells could express kallistatin after plasmid transfection. The proliferation and migration of human umbilical vein endothelial cells were inhibited, but the apoptosis rate was not affected. The proliferation rates of all the three tested lung cancer cells, such as NCI-H446, NCI-H460 and A549, were inhibited, and their apoptosis rates were enhanced, but different cells behaved differently. In subcutaneous NCI-H446 xenograft model, intratumor injection of pKal inhibited the growth of lung cancer by reducing angiogenesis and proliferation of tumor cells. In conclusion, this study demonstrated the efficacy of plasmid-mediated expression of kallistatin to lung cancer related cells, thus providing a basis for their clinical application in the treatment of lung cancer.
Subject(s)
Animals , Humans , Male , Mice , Antigens, CD34 , Metabolism , Apoptosis , Cadherins , Metabolism , Cell Line, Tumor , Cell Movement , Cell Proliferation , Cells, Cultured , Human Umbilical Vein Endothelial Cells , Cell Biology , Metabolism , Ki-67 Antigen , Metabolism , Lung Neoplasms , Metabolism , Pathology , Mice, Inbred BALB C , Mice, Nude , Neoplasm Transplantation , Neovascularization, Pathologic , Plasmids , Serpins , Genetics , Metabolism , Physiology , Transfection , Tumor BurdenABSTRACT
<p><b>BACKGROUND</b>Molecular hydrogen, as a novel antioxidant, has been proven effective in treating many diseases. This study aimed to evaluate the therapeutic effects of hydrogen saturated saline in treatment of a rat model of diabetes mellitus and a rat model of insulin resistant.</p><p><b>METHODS</b>A rat diabetes mellitus model was established by feeding a high fat/high carbohydrate diet followed by injection of a small dose of streptozotocin, and an insulin resistant model was induced with a high glucose and high fat diet. Hydrogen saturated saline was administered to rats with both models conditions on a daily basis for eight weeks. A pioglitazone-treated group and normal saline-treated group served as positive and negative controls. The general condition, body weight, blood glucose, blood lipids, and serum insulin levels of rats were examined at the 8th week after treatment. The oxidative stress indices, including serum superoxide dismutase (SOD), glutathione (GSH) and malondialdehyde (MDA) were also evaluated after eight weeks of treatment using the commercial kits.</p><p><b>RESULTS</b>Hydrogen saturated saline showed great efficiency in improving the insulin sensitivity and lowering blood glucose and lipids. Meanwhile, the therapeutic effects of hydrogen saturated saline were superior to those of pioglitazone. Hydrogen saturated saline markedly attenuated the MDA level and elevated the levels of antioxidants SOD and GSH.</p><p><b>CONCLUSION</b>Hydrogen saturated saline may improve the insulin resistance and alleviate the symptoms of diabetes mellitus by reducing the oxidative stress and enhancing the anti-oxidant system.</p>
Subject(s)
Animals , Rats , Diabetes Mellitus, Experimental , Drug Therapy , Hydrogen , Therapeutic Uses , Hypoglycemic Agents , Therapeutic Uses , Insulin Resistance , Oxidative Stress , Sodium Chloride , Chemistry , Thiazolidinediones , Therapeutic UsesABSTRACT
<p><b>OBJECTIVE</b>To investigate the expression of casein kinase 2β (ck2β) in colorectal cancer in relation to the metastatic ability of the cancer cells.</p><p><b>METHODS</b>The expression of ck2β in 46 normal colorectal mucosa, 20 colorectal adenomas and 66 colorectal cancers were detected immunohistochemically. In colorectal cancer cells, Ck2β protein expression was knockdown by RNA interference using ck2β-specific small interfering RNA (siRNA) and the interference efficiency was assessed by Western blotting. The effect of ck2β gene knockdown on the proliferation of the colorectal cancer cells was tested with colony formation assay, and the changes in the invasive ability of the cells were observed using Transwell chamber assay.</p><p><b>RESULTS</b>Negative or weak ck2β expression was detected in normal colorectal mucosa, with nuclear positivity in 8.7% and cytoplasmic positivity in 13.0% of the cases. Colorectal adenomas showed moderate ck2β expression, with 60% cases showing positivity in the cell nuclei and 40% in the cytoplasm. In colorectal cancers, significantly stronger expression of ck2β was found than that in colorectal adenomas and normal colorectal mucosa (P<0.05), and 75.8% cases showed positivity in the cell nuclei and 62.1% showed cytoplasmic positivity; the expression of ck2β protein in colorectal cancers with lymph node metastasis was even higher (P<0.05). Ck2β knockdown obviously inhibited the proliferation and invasiveness of colorectal cancer cells in vitro.</p><p><b>CONCLUSION</b>The high expression of ck2β in colorectal cancer is closely correlated to the carcinogenesis and metastasis of the tumor.</p>
Subject(s)
Adult , Aged , Female , Humans , Male , Middle Aged , Young Adult , Casein Kinase II , Metabolism , Cell Proliferation , Cell Transformation, Neoplastic , Colorectal Neoplasms , Metabolism , Pathology , Intestinal Mucosa , Metabolism , Pathology , Tumor Cells, CulturedABSTRACT
The oral administration of bioactive macromolecular drugs such as proteins, peptides and nucleic acids represents unprecedented challenges from the drug delivery point of view. One key consideration is how to overcome the gastrointestinal tract absorption barrier. Recent studies suggest that microfold cell (M cell), a kind of specialized antigen-sampling epithelial cell which is characterized by a high endocytic rate and low degradation ability, may play an important role in macromolecule oral absorption. The development of an in vitro M cell coculture system and its modified models greatly advanced the study of M cells and the development of oral delivery system for macromolecular drugs. The special structure, function and formation characteristics, and biomarkers of M cell are summarized in this review. The applications of in vitro M cell models in developing oral delivery system ofbioactive macromolecular drugs are discussed.
Subject(s)
Animals , Humans , Administration, Oral , Drug Delivery Systems , Methods , Intestinal Mucosa , Cell Biology , Macromolecular Substances , Pharmacokinetics , Models, Biological , Peptides , Pharmacokinetics , Peyer's Patches , Cell Biology , Proteins , Pharmacokinetics , Vaccines , PharmacokineticsABSTRACT
The oral administration of bioactive macromolecular drugs such as proteins, peptides and nucleic acids represents unprecedented challenges from the drug delivery point of view. One key consideration is how to overcome the gastrointestinal tract absorption barrier. Recent studies suggest that microfold cell (M cell), a kind of specialized antigen-sampling epithelial cell which is characterized by a high endocytic rate and low degradation ability, may play an important role in macromolecule oral absorption. The development of an in vitro M cell coculture system and its modified models greatly advanced the study of M cells and the development of oral delivery system for macromolecular drugs. The special structure, function and formation characteristics, and biomarkers of M cell are summarized in this review. The applications of in vitro M cell models in developing oral delivery system ofbioactive macromolecular drugs are discussed.
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<p><b>OBJECTIVE</b>To investigate the effect of compound Danshen Dripping Pill (DSP) on carotid arterial intima-media thickness (IMT) in patients with type 2 diabetes mellitus (T2DM).</p><p><b>METHODS</b>One hundred and thirty T2DM patients were assigned to four groups, 32 in the Group A, the control group treated with blood glucose (BG) and blood pressure (BP) controlling; 32 in the Group B, with BG, BP and blood lipid (BL) controlling, 32 in Group C with BG, BP, BL controlling and vitamin E administration, and 34 in Group D with BG, BP, BL controlling and DSP administration. Patients in Group D were subdivided by Chinese medicine syndrome differentiation into four types, 8 of Yin-deficiency with flourishing heat type (YDFH), 5 of both qi-yin deficient type (BQYD), 8 of both yin-yang deficient type (BYYD) and 13 of blood-stasis and qi-stagnant type (BSQS). Fasting blood glucose (FBG), BP and BL in patients were observed periodically, and IMT in them were measured by ultrasonography before treatment, as well as at the end of the 1st, 3rd, and 5th year of treatment to dynamically observe the changes of IMT and condition of plaque formation, and analyze the relation between them with FBG, BP and BL.</p><p><b>RESULTS</b>The 5-year follow-up was performed in 105 patients. In the observation period, level of total cholesterol (TC) showed a decreasing trend and level of high density cholesterol (HDL-C) showed an increasing trend in all the 4 groups, the improvements in Group C and D were slightly better than those in Group B, while significantly superior to those in Group A; the changes of FBG and glycosylated hemoglobin (HbAlc) were insignificant in the 4 groups. IMT and numbers of atheroma plaque increased gradually in all groups in the observation period, however, the changes in Group D were lesser than those in other groups, showing significant difference (P < 0.01). It was showed that the increasing of cervical carotid IMT in T2DM patients was correlated with levels of HbAlc, HDL-C, LDL-C, triglyceride and TC, especially in Group D.</p><p><b>CONCLUSION</b>DSP might delay the occurrence and development of diabetic macro-vascular disease.</p>
Subject(s)
Female , Humans , Male , Middle Aged , Carotid Arteries , Pathology , Carotid Intima-Media Thickness , Diabetes Mellitus, Type 2 , Drug Therapy , Pathology , Diabetic Angiopathies , Pathology , Diagnosis, Differential , Drugs, Chinese Herbal , Therapeutic Uses , Medicine, Chinese Traditional , Phytotherapy , Salvia miltiorrhiza , Chemistry , Tunica Intima , Pathology , Tunica Media , PathologyABSTRACT
<p><b>BACKGROUND</b>There is no agreement as to whether intensive glucose control in type 2 diabetes can reduce the incidence of macrovascular events in these patients. We performed a meta-analysis comparing intensive glucose control or conventional glucose control in randomized controlled trials.</p><p><b>METHODS</b>Databases including MEDLINE, EMBASE, and Cochrane controlled trials register, the Cochrane Library, and Science Citation Index were searched to find relevant trials. Outcome measures were the incidence of major macrovascular events.</p><p><b>RESULTS</b>Six trials involving 28 065 patients were included. Analysis suggested that there was an obviously decreased incidence of major macrovascular events in patients having intensive glucose treatment vs. controls (RR 0.92; 95%CI 0.87, 0.98; P = 0.005). However, intensive glycemia control strategies in type 2 diabetes showed no significant impact on the incidence of death from any cause compared with conventional glycemia control strategies, intensive 14.7%, controls 12.0% (RR 0.95; 95%CI 0.80, 1.12; P = 0.55), as well as on the incidence of cardiovascular death, intensive 3.7%, controls 3.6% (RR 1.10, 95%CI 0.79, 1.53; P = 0.57).</p><p><b>CONCLUSIONS</b>Control of glycemia to normal (or near normal levels) in type 2 diabetes appears to be effective in reducing the incidence of major macrovascular events, but there were no significant differences of either the mortality from any cause or from cardiovascular death between the two glycemia-control strategies.</p>
Subject(s)
Humans , Blood Glucose , Diabetes Mellitus, Type 2 , Blood , Drug Therapy , Mortality , Diabetic Angiopathies , Glycated Hemoglobin , Randomized Controlled Trials as TopicABSTRACT
2-Amino-3-hydroxylpyridinE-H2O2-horseradish peroxidase (HRP) voltammetric enzymE-linked immunoassay based on N-heterocyclic substrate has been successfully applied for the detection of carcionembryonic antigen(CEA) in human serum. 2-Amino-3-hydroxylpyridine is oxidized with H2O2 catalyzed by HRP, and the resulting electroactive product produces a sensitive voltammetric peak at the potential of 0.36 V(vs. SCE) in Britton-Robinson (B-R) buffer solution. By this voltammetric peak, free HRP can be measured and immunoassay of HRP label can be developed. The enzymE-catalyzed reaction conditions and voltammetric detection conditions have been optimized, and the electrode procedure of the enzymatic product was investigated. The selected optimum reaction conditions were that the reaction medium was pH 6.0 B-R buffer solution for 10 mL reaction solution containing 1.0 mL of 0.2 mol/L B-R buffer solution, 3.0 mL of 8.0 mmol/L 2-amino-3-hydroxylpyridine solution and 1.5 mL of 0.5 mmol/L H2O2 solution, and the reaction time was 30 min at 37 ℃. The optimum detection conditions were that the supporting electrolyte was pH 7.0 B-R buffer solution for 10 mL of the overall detection solution containing 5 mL of reaction solution and 1.0 mL of 0.2 mol/L B-R buffer solution. The optimum instrumental conditions for the detection were chosen as follows: the initial potential, 0.00 V; the final potential, -0.80 V; the potential scanning rate, 400 mV/s; the mercury drop standing time, 7 s. Under the optimum conditions, the linear range for detection of free HRP was 4.0×10-4-1.0 μg/L with a detection limit of 0.12 μg/L. Based on the new immunoassay system, the linear range of the detection to CEA was 0.50-80 μg/L and the detection limit was 0.50 μg/L, which is 10 times lower than that of traditional spectrophotometric enzymE-linked immunosorbent assay method. The 2-Amino-3-hydroxylpyridinE-H2O2-HRP voltammetric enzymE-linked immunoassay new system exhibits excellent performance having wider linear range and lower detection limit. The new method is inexpensive and simple. It has a great potential in clinical diagnosis.
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<p><b>OBJECTIVE</b>To investigate the effect of protein kinase CK2 gene silencing on the radiosensitization in human nasopharyngeal carcinoma (NPC) cells and its possible mechanism.</p><p><b>METHODS</b>RNA interference (RNAi) technique was used to down-regulate the protein kinase CK2alpha expression in 5-8F cells, and clonogenic assay was employed to observe the changes in the radiosensitivity of the cells. DNA double-strand break was assessed by immunofluorescence staining of gamma-H2AX foci, and the cell apoptosis was examined using Annexin V-FITC/PI double-staining flow cytometry.</p><p><b>RESULTS</b>CK2alpha protein was successfully silenced by siRNA. CK2alpha knockdown significantly decreased the clonogenic activity and increased the radiosensitivity of the NPC cells. After a 15-min exposure of the cells to 1 Gy radiation, significant difference occurred in the gamma-H2AX foci between CK2alpha knockdown cells and the control cells (P<0.01). CK2alpha silencing significantly increased the cell apoptosis after the exposure (P<0.01).</p><p><b>CONCLUSIONS</b>Protein kinase CK2 plays an important role in the radiosensitivity of the NPC cells, and suppression of its expression might be a potential therapeutic approach of cancer.</p>
Subject(s)
Animals , Humans , Annexin A5 , Metabolism , Casein Kinase II , Genetics , Metabolism , Cell Line, Tumor , Histones , Genetics , Nasopharyngeal Neoplasms , Genetics , Pathology , RNA Interference , RNA, Small Interfering , Genetics , Radiation Tolerance , Genetics , TransfectionABSTRACT
<p><b>OBJECTIVE</b>To investigate the prevalence of peripheral arterial disease (PAD) in China type 2 diabetic patients and to demonstrate the relationships between putative risk factors and PAD.</p><p><b>METHODS</b>In total 1,397 type 2 diabetic patients aged 50 years and older were enrolled and determined ankle-brachial index (ABI) and brachial-ankle pulse wave velocity (baPWV) in 15 Class III Grade A hospitals in 7 major cities of China.</p><p><b>RESULTS</b>Mean patient age was 63.7 +/- 8.2 years and mean duration of diabetes mellitus was 9.39 +/- 7.4 years. Two hundreds and seventy-two (19.47%) patients were diagnosed as PAD by ABI < 0.9, 122 (18.37%) in male and 150 (20.46%) in female. PAD patients had a significantly longer duration of diabetes mellitus, higher hemoglobin A1c, and a significantly lower mean body mass index than non-PAD ones. Aging, smoking, and systolic blood pressure were found to be positively related with the prevalence of PAD. In terms of lipid profiles, no variable was found to relate with PAD. Notably, baPWV showed as the same significant guiding index for PAD, almost matched with ABI.</p><p><b>CONCLUSIONS</b>PAD is a common complication in China type 2 diabetic patients. Therefore, PAD screening and treatment should be emphasized for diabetic patients with high risk factors.</p>
Subject(s)
Aged , Female , Humans , Male , Middle Aged , China , Epidemiology , Diabetic Angiopathies , Epidemiology , Peripheral Vascular Diseases , Epidemiology , Prevalence , Risk Factors , Urban PopulationABSTRACT
<p><b>OBJECTIVE</b>To compare the immunity of morbid obesity (MO) before and after laparoscopic adjustable gastric banding (LAGB).</p><p><b>METHODS</b>15 cases, with a mean body mass index (BMI) of 35.8 kg/m(2), were treated by LAGB from Jun. 2003 to Oct. 2003 in our department. Patients' immune parameters were determined preoperatively and 1, 3 and 6 months postoperatively. 15 cases with a normal BMI (23.6 kg/m(2)) were set as controls.</p><p><b>RESULTS</b>Before surgery, the MO had a significant lower level of CD(4)(+), CD(4)(+)/CD(8)(+) and a higher level of serum interleukin-2 (IL-2), Interleukin-6 (IL-6) than the controls (P < 0.01). There was a significant reduction of weight and BMI 6 months postoperatively (P < 0.01). At the same time, CD(4)(+) increased and serum IL-2 decreased significantly. But CD(4)(+)/CD(8)(+)and serum IL-2, IL-6 were still abnormal compare to the controls.</p><p><b>CONCLUSIONS</b>MO may combined with an abnormal immunity. But after enough weight loss induced by LAGB, it can be partly reversed.</p>
Subject(s)
Adolescent , Adult , Female , Humans , Male , Follow-Up Studies , Gastroplasty , Methods , Laparoscopy , Obesity, Morbid , Allergy and Immunology , General Surgery , Weight LossABSTRACT
<p><b>OBJECTIVE</b>Increased vascular endothelial growth factor (VEGF) and VEGF receptor expression is the important biological response under shear stress, ischemia and hypoxia conditions. Mechanical vibration induced cochlea shear stress and trauma obviously upregulate VEGF and VEGF receptor 2 (VEGFR2) expression in the cochlea. To evaluate the possibility of VEGF varying the transport in blood-labyrinth barrier and blood-perilymphatic barrier.</p><p><b>METHODS</b>Eleven guinea pigs, male and female, weighing from 300 g to 900 g were kept under general anaesthesia with xylazine (16 mg/kg) and ketamine (60 mg/kg) for both drug delivery and MRI measurement. VEGF (6 ears) and phosphate-buffered saline (PBS, 5 ears) were delivered to the inner ear via the round window membrane (soaked in gelfoam). The T1 contrast agent gadodiamide (Gd-DTPA-BMA) chelated bound paramagnetic gadolinium was used as the inner ear barrier transportation tracer. A Bruker Biospec Avance 47/40 experimental MRI system with a magnetic field strength of 4. 7 Tesla and a 40 cm bore was used for the 2-dimensional cochlea MRI evaluation. The Paravision software was used for image intensity measurement and the Adobe Photoshop 6.0 software was used for image presentation.</p><p><b>RESULTS</b>VEGF induced significant Gd uptake in the scala tympani and scala vestibuli, but had little effect on the uptake of Gd in the scala media.</p><p><b>CONCLUSIONS</b>VEGF significantly increased the transportation of blood-perilymphatic barrier and adapted the inner ear for compensation and repair.</p>
Subject(s)
Animals , Female , Male , Blood-Brain Barrier , Blood-Retinal Barrier , Ear, Inner , Metabolism , Guinea Pigs , Magnetic Resonance Imaging , Vascular Endothelial Growth Factor A , PharmacologyABSTRACT
Bio-heat transfer in the tongue body is studied combining the mechanism of tongue inspection in Traditional Chinese Medicine. Parameters such as the temperature of lingual surface, the blood perfusion of the dog's tongue and so on are measured and the influences of the blood perfusion, the arterial blood temperature, the arterial cross-sectional areas and positions on the temperature distribution in the cross-sections are studied. Then the two-dimensional temperature fields in different cross-sections are numerically solved by the finite element method (FEM). The results show that the vascular cross-sectional areas vary with the elasticity change of the vessel walls resulting from the variation of the blood perfusion. And the temperature distribution in the cross-section mainly depends on the arterial cross-sectional areas and positions. The results can help to analyze the bio-heat transfer characteristic of the tongue. According to this method, the sectional temperature fields in whatever place of the tongue can be calculated under different blood perfusion and on the condition that the influence of the venous blood temperature on the temperature field of the tongue is considered. We can further probe into the relationship between the temperature fields of the lingual surface and that of cross-sections. This can provide the foundation for further investigation into the bio-heat transfer mechanism and the calculation on three-dimensional temperature fields of the tongue.
Subject(s)
Animals , Dogs , Body Temperature , Physiology , Finite Element Analysis , In Vitro Techniques , Models, Biological , Regional Blood Flow , Thermal Conductivity , Tongue , PhysiologyABSTRACT
Objective To investigate the expression of protein tyrosine phosphatase(PTP)1B in pancreas of obese rats induced by high fat diet.Methods Twenty male SD rats were randomly divided into control group on regular diet(n=10),and obesity model group on high fat diet(n=10).After twelve weeks,fasting serum insulin,blood glucose,triglyceride,total cholesterol,epididymal fat weight were measured and the histomorphological change in liver were observed;glucose tolerance test and insulin releasing test were performed; The protein expression of PTP-1B in pancreas of rats was determined with Western blotting and immunohistochemistry.The phosphorylation of insulin receptor substrate-1(IRS-1)and insulin receptor(IR)were detected by immuno-precipitation.Results(1)The insulin sensitive index was significantly decreased in the high-fat-diet rats compared with the normal rats(0.36?0.18 vs 0.91?0.28,P