Human HDLs have highly heterogeneous composition. Plasma concentrations of HDL with apoC-III and of apoE in HDL predict higher incidence of coronary heart disease (CHD). The concentrations of HDL-apoA-I containing apoE, apoC-III, or both and their distribution across HDL sizes are unknown. We studied 20 normal weight and 20 obese subjects matched by age, gender, and race. Plasma HDL was separated by sequential immunoaffinity chromatography (anti-apoA-I, anti-apoC-III, anti-apoE), followed by nondenaturing-gel electrophoresis. Mean HDL-cholesterol concentrations in normal weight and obese subjects were 65 and 50 mg/dl (P = 0.009), and total apoA-I concentrations were 119 and 118 mg/dl, respectively. HDL without apoE or apoC-III was the most prevalent HDL type representing 89% of apoA-I concentration in normal weight and 77% in obese (P = 0.01) individuals; HDL with apoE-only was 5% versus 8% (P = 0.1); HDL with apoC-III-only was 4% versus 10% (P = 0.009); and HDL with apoE and apoC-III was 1.5% versus 4.6% (P = 0.004). Concentrations of apoE and apoC-III in HDL were 1.5–2× higher in obese subjects (P ≤ 0.004). HDL with apoE or apoC-III occurred in all sizes among groups. Obese subjects had higher prevalence of HDL containing apoE or apoC-III, subfractions associated with CHD, whereas normal weight subjects had higher prevalence of HDL without apoE or apoC-III, subfractions with protective association against CHD. Human HDLs have highly heterogeneous composition. Plasma concentrations of HDL with apoC-III and of apoE in HDL predict higher incidence of coronary heart disease (CHD). The concentrations of HDL-apoA-I containing apoE, apoC-III, or both and their distribution across HDL sizes are unknown. We studied 20 normal weight and 20 obese subjects matched by age, gender, and race. Plasma HDL was separated by sequential immunoaffinity chromatography (anti-apoA-I, anti-apoC-III, anti-apoE), followed by nondenaturing-gel electrophoresis. Mean HDL-cholesterol concentrations in normal weight and obese subjects were 65 and 50 mg/dl (P = 0.009), and total apoA-I concentrations were 119 and 118 mg/dl, respectively. HDL without apoE or apoC-III was the most prevalent HDL type representing 89% of apoA-I concentration in normal weight and 77% in obese (P = 0.01) individuals; HDL with apoE-only was 5% versus 8% (P = 0.1); HDL with apoC-III-only was 4% versus 10% (P = 0.009); and HDL with apoE and apoC-III was 1.5% versus 4.6% (P = 0.004). Concentrations of apoE and apoC-III in HDL were 1.5–2× higher in obese subjects (P ≤ 0.004). HDL with apoE or apoC-III occurred in all sizes among groups. Obese subjects had higher prevalence of HDL containing apoE or apoC-III, subfractions associated with CHD, whereas normal weight subjects had higher prevalence of HDL without apoE or apoC-III, subfractions with protective association against CHD. Epidemiological studies have shown that low plasma levels of HDL cholesterol (HDL-C) are strongly associated with an elevated risk of coronary heart disease (CHD) (1Gordon T. Castelli W.P. Hjortland M.C. Kannel W.B. Dawber T.R. High density lipoprotein as a protective factor against coronary heart disease: the Framingham Study.Am. J. Med. 1977; 62: 707-714Abstract Full Text PDF PubMed Scopus (4087) Google Scholar, 2.SharrettA. R.BallantyneC. M.CoadyS. A.HeissG.SorlieP. D.CatellierD.PatschW.; Atherosclerosis Risk in Communities Study Group. 2001. 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VLDL, apoliproteins B, CIII, and E, and risk of recurrent coronary events in the Cholesterol and Recurrent Events (CARE) Trial.Circulation. 2000; 102: 1886-1892Crossref PubMed Scopus (412) Google Scholar, 16Vaisar T. Mayer P. Nilsson E. Zhao X.Q. Knopp R. Prazen B.J. HDL in humans with cardiovascular disease exhibits a proteomic signature.Clin. Chim. Acta. 2010; 411: 972-979Crossref PubMed Scopus (68) Google Scholar, 17Jensen M.K. Rimm E.B. Furtado J.D. Sacks F.M. Apolipoprotein C-III as a potential modulator of the association between HDL-cholesterol and incident coronary heart disease.J. Am. Heart Assoc. 2012; 1jah3-e000232Crossref PubMed Google Scholar). ApoE and apoC-III are found on the surface of both triglyceride-rich lipoproteins (TRLs) and HDL (18Campos H. Perlov D. Khoo C. Sacks F.M. Distinct patterns of lipoproteins with apoB defined by presence of apoE or apo C-III in hypercholesterolemia and hypertriglyceridemia.J. 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More recently, a proteomic analysis showed greater apoE enrichment in small-size HDL (HDL3) in subjects with established coronary artery disease than in normal controls (16Vaisar T. Mayer P. Nilsson E. Zhao X.Q. Knopp R. Prazen B.J. HDL in humans with cardiovascular disease exhibits a proteomic signature.Clin. Chim. Acta. 2010; 411: 972-979Crossref PubMed Scopus (68) Google Scholar). Similarly, HDL containing apoC-III independently predicts increased risk of an initial coronary event in separate cohorts of men and women (17Jensen M.K. Rimm E.B. Furtado J.D. Sacks F.M. Apolipoprotein C-III as a potential modulator of the association between HDL-cholesterol and incident coronary heart disease.J. Am. Heart Assoc. 2012; 1jah3-e000232Crossref PubMed Google Scholar), and a high ratio of apoC-III to apoA-I in HDL predicts recurrent coronary events (15Sacks F.M. Alaupovic P. Moye L.E. Cole T.G. Sussex B. Stampfer M.J. Pfeffer M.A. Braunwald E. VLDL, apoliproteins B, CIII, and E, and risk of recurrent coronary events in the Cholesterol and Recurrent Events (CARE) Trial.Circulation. 2000; 102: 1886-1892Crossref PubMed Scopus (412) Google Scholar). Distinct HDL speciation based on apoC-III and apoE content is not firmly established in the literature. ApoC-III and apoE long have been known to be present in HDL (19Alaupovic P. Significance of apolipoproteins for structure, function, and classification of plasma lipoproteins.Methods Enzymol. 1996; 263: 32-60Crossref PubMed Google Scholar, 20Shah A.S. Tan L. Long J.L. Davidson W.S. Proteomic diversity of high-density lipoproteins: our emerging understanding of its importance in lipid transport and beyond.J. Lipid Res. 2013; 54: 2575-2585Abstract Full Text Full Text PDF PubMed Scopus (265) Google Scholar). Findings in proteomics studies showed that apoC-III and apoE are present throughout the range of sizes of HDL that are prepared by ultracentrifugation or gel filtration, or by precipitation of apoB lipoproteins (21Davidson W.S. Silva R.A.G.D. Chantepie S. Lagor W.R. Chapman M.J. Kontush A. Proteomic analysis of defined HDL subpopulations reveals particle-specific protein clusters: relevance to antioxidative function.Arterioscler. Thromb. Vasc. Biol. 2009; 29: 870-876Crossref PubMed Scopus (327) Google Scholar, 22Gordon S.M. Deng J. Lu L.J. Davidson W.S. Proteomic characterization of human plasma high density lipoprotein fractionated by gel filtration chromatography.J. Proteome Res. 2010; 9: 5239-5249Crossref PubMed Scopus (170) Google Scholar, 23Orsoni A. Saheb S. Levels J. Dallinga-Thie G. Atassi M. Bittar R. Robillard P. Bruckert E. Kontush A. Carrie A. et al.LDL-apheresis depletes apoE-HDL and pre-b1-HDL in familial hypercholesterolemia: relevance to atheroprotection.J. 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Apolipoprotein C-III and the metabolic basis for hypertriglyceridemia and the dense low-density lipoprotein phenotype.Circulation. 2010; 121: 1722-1734Crossref PubMed Scopus (181) Google Scholar); whether apoC-III and apoE coexist on HDL as they do on VLDL and LDL (18Campos H. Perlov D. Khoo C. Sacks F.M. Distinct patterns of lipoproteins with apoB defined by presence of apoE or apo C-III in hypercholesterolemia and hypertriglyceridemia.J. Lipid Res. 2001; 42: 1239-1249Abstract Full Text Full Text PDF PubMed Google Scholar, 19Alaupovic P. Significance of apolipoproteins for structure, function, and classification of plasma lipoproteins.Methods Enzymol. 1996; 263: 32-60Crossref PubMed Google Scholar, 24Zheng C. Khoo C. Furtado J. Sacks F.M. Apolipoprotein C-III and the metabolic basis for hypertriglyceridemia and the dense low-density lipoprotein phenotype.Circulation. 2010; 121: 1722-1734Crossref PubMed Scopus (181) Google Scholar, 25.MendivilC. O.RimmE. B.FurtadoJ.SacksF. M.. 2013. 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Even so, there remains the issue of conflicting findings involving previous studies proposing that most if not all apoE in HDL is found on particles that do not have apoA-I as suggested by nonoverlapping regions of apoE and apoA-I on nondenaturing two-dimensional gel electrophoresis with immunoblotting (27Asztalos B.F. Schaefer E.J. Horvath K.V. Yamashita S. Miller M. Franceschini G. Calabresi L. Role of LCAT in HDL remodeling: investigation of LCAT deficiency states.J. Lipid Res. 2007; 48: 592-599Abstract Full Text Full Text PDF PubMed Scopus (135) Google Scholar, 28Krimbou L. Marcil M. Chiba H. Genest Jr, J. Structural and functional properties of human plasma high density-sized lipoprotein containing only apoE particles.J. Lipid Res. 2003; 44: 884-892Abstract Full Text Full Text PDF PubMed Scopus (46) Google Scholar) or with reconstituted HDL particles (29Rye K.A. Bright R. Psaltis M. Barter P.J. 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The purpose of this study was to examine apoA-I-containing HDL subpopulations as defined by apoE and apoC-III content as well as by size in the context of normal body weight and obesity, a clinical condition related to CHD. We studied the distribution of these different apolipoprotein-defined HDL types in a group with normal HDL-C levels and normal body weight and compared it with that of an obese group, a common high-risk phenotype associated with low HDL-C. Elevated apoC-III concentrations in HDL (34Chan D.C. Nguyen M.N. Watts G.F. Barrett P.H. Apolipoprotein C-III transport in centrally obese men: associations with very low-density lipoprotein apolipoprotein B and high-density lipoprotein apolipoprotein A-I metabolism.J. Clin. Endocrinol. Metab. 2008; 93: 557-564Crossref PubMed Scopus (57) Google Scholar, 35Onat A. Hergenç G. Ayhan E. U ur M. Kaya H. Tuncer M. Can G. Serum apolipoprotein C-III in high-density lipoprotein: a key diabetogenic risk factor in Turks.Diabet. Med. 2009; 26: 981-988Crossref PubMed Scopus (52) Google Scholar), as well as increased apoC-III production (36Cohn J.S. Tremblay M. Batal R. Jacques H. Rodriguez C. Steiner G. Mamer O. Davignon J. Increased apoC-III production is a characteristic feature of patients with hypertriglyceridemia.Atherosclerosis. 2004; 177: 137-145Abstract Full Text Full Text PDF PubMed Scopus (57) Google Scholar), are common features of obesity. We hypothesized that the distribution of HDL particles is disturbed with obesity, such that obese subjects have higher concentrations of HDL with apoC-III as well as HDL with apoE, and lower concentrations of HDL without apoE or apoC-III contributing to their increased risk for CHD. The study population consisted of 40 participants, 26 women and 14 men, age 30–67 years, assigned to two different groups based on BMI. The obese group (n = 20) was defined as having a BMI ≥30, while the normal weight group (n = 20) was defined as having a BMI ≤25. Subjects for the obese group were selected from the POUNDS LOST study, a randomized clinical trial comparing diets for weight loss (37Sacks F.M. Bray G.A. Carey V.J. Smith S.R. Ryan D.H. Anton S.D. McManus K. Champagne C.M. Bishop L.M. Laranjo N. et al.Comparison of weight-loss diets with different compositions of fat, protein, and carbohydrates.N. Engl. J. Med. 2009; 360: 859-873Crossref PubMed Scopus (1374) Google Scholar). Briefly, the eligibility criteria for the parent study were age 30–70 years and BMI between 25 kg/m2 and ≤40 kg/m2. Major exclusions in the parent study were diabetes, unstable cardiovascular disease, and use of lipid-lowering medications or medications that affect body weight. Participants selected from the POUNDS LOST study for our obese group had a BMI ≥30, and the samples used were baseline samples, prior to dietary intervention. Participants for the normal weight group were healthy volunteers with a BMI ≤25 and were selected using the same major exclusions as for the obese group. Obese subjects were matched to normal weight subjects by age, gender, and race. We aimed to study a representative group of matched normal and obese participants that would have more generalizability than a select population subgroup. All study participants gave informed consent, and the Human Subjects Committees of the Harvard School of Public Health and Brigham and Women's Hospital approved the study. Blood was collected from participants following at least an 8 h fast. Serum was separated and aliquotted by trained personnel at each clinical site and stored at −80°C. Vials containing 0.5 ml of frozen plasma were sent from Pennington Biomedical Research Center to the lipoprotein laboratory at the Harvard School of Public Health. Samples from both study groups were collected and stored in frozen vials for an average of 2 years before analysis of lipoprotein types was conducted, with the exception of five samples from our normal weight study group that were collected and stored in frozen vials 6 months before analysis. The laboratory personnel were blinded to the samples' group status. Plasma samples were removed from cryogenic storage, thawed, and filtered for the removal of fibrinogen and coagulation products using Pall (R) Acrodisc filters (5 µm). ApoA-I-containing lipoproteins were then separated from plasma using immunoaffinity-purified polyclonal anti-apoA-I antibodies as follows (Fig. 1): 1 ml of filtered plasma was loaded and incubated in 20 ml Econo-Pac columns (Bio-Rad Laboratories, Hercules, CA) packed with 2.5 ml of affinity-purified polyclonal goat anti-human apoA-I antibodies (Academy Bio-Medical Co., Houston, TX) bound to Sepharose 4B resin. The unbound lipoproteins were collected by gravity flow followed by washes with PBS and stored at −80°C. The bound fraction, which was the fraction of interest, was then eluted from the columns with 3M sodium thiocyanate (NaSCN) in PBS and immediately desalted by multiple rinses in Vivaspin 20 ultrafiltration centrifugal device with polyethersulfone (PES) membrane at 10,000 Da molecular weight cutoff (MWCO) (Sartorius Stedim Biotech, Germany), ending with a final sample volume of 500 μl. This apoA-I-containing fraction was then loaded sequentially onto anti-apoC-III columns followed by anti-apoE columns for further separation of apoA-I-containing lipoproteins by apoE and apoC-III content (18Campos H. Perlov D. Khoo C. Sacks F.M. Distinct patterns of lipoproteins with apoB defined by presence of apoE or apo C-III in hypercholesterolemia and hypertriglyceridemia.J. Lipid Res. 2001; 42: 1239-1249Abstract Full Text Full Text PDF PubMed Google Scholar, 38Khoo C. Judge H. Sacks F.M. Effects of estrogenic oral contraceptives on the lipoprotein B particle system defined by apolipoproteins E and C-III content.J. Lipid Res. 1999; 40: 202-212Abstract Full Text Full Text PDF PubMed Google Scholar). In detail, the apoA-I-containing fraction was loaded and incubated under similar conditions onto columns packed with affinity-purified polyclonal antibodies anti-apoC-III (DMA, Arlington, TX). The unbound fractions (C-IIIˉ) were collected by gravity flow from the columns, and the resin was washed with PBS. The bound fraction (C-III+) was eluted by incubation with 3M NaSCN in PBS and immediately desalted by multiple rinses as described for the apoA-I-containing fraction. The C-III− fractions and the dialyzed C-III+ fractions were then finally loaded and incubated in columns packed with affinity-purified polyclonal antibodies anti-apoE (Genzyme, Cambridge, MA). The same elution protocol used for the anti-apoA-I and the anti-apoC-III resin was carried out. This yielded four distinct subfractions: HDL without apoE or apoC-III (E−CIII−), HDL with apoE but without apoC-III (E+CIII−), HDL with apoC-III but without apoE (E−CIII+), and HDL with both apoE and apoC-III (E+CIII+). All incubations occurred overnight in the cold room with constant mixing. The efficiencies of the apoC-III and apoE immunoaffinity separation (percentage of ligand removed from plasma by the resin) were 97% and 94%, respectively. After collection of the bound fraction, the immunoaffinity columns were washed three times with NaSCN and with PBS containing EDTA. Then the columns were reconditioned by 0.1 M acetic acid and finally washed with PBS. The columns were completely clean of apos after this procedure. For example, the third wash with NaSCN eluted <0.4% of plasma apoA-I. No apoA-I was detected after a fourth wash, showing that the wash procedure removed all the bound apoA-I. To address a concern on the possible transfer of apoC-III from VLDL to HDL during freezing or storage (39Cohn J.S. Rodriguez C. Jacques H. Tremblay M. Davignon J. Storage of human plasma samples leads to alterations in the lipoprotein distribution of apoC-III and apoE.J. Lipid Res. 2004; 45: 1572-1579Abstract Full Text Full Text PDF PubMed Scopus (22) Google Scholar), we measured apoC-III in VLDL, LDL, and HDL immediately after plasma separation by density gradient ultracentrifugation and after 5 months of storage at −80°C in samples from four normal individuals. The distribution of apoC-III between TRLs and HDL was similar [14% in VLDL, 16% in LDL, and 70% in HDL, when analyzed fresh; 12% in VLDL, 15% in LDL, and 73% in HDL after frozen storage; multivariate ANOVA (MANOVA) P = 0.1]. We then tested the effects of storage and thawing on apoA-I-containing HDL when separated by immunoaffinity chromatography and found no differences in apoE or apoC-III distribution on HDL between fresh and previously stored control plasma samples: mean apoA-I HDL without apoE was 135 ± 5 mg/dl and HDL with apoE 10 ± 0.7 mg/dl in the fresh samples versus 134 ± 5.6 mg/dl and 7 ± 2 mg/dl in the frozen samples (P = 0.3 and 0.1); similarly, mean apoA-I HDL without apoC-III was 137 ± 5 mg/dl and HDL with apoC-III 8.4 ± 1 mg/dl in the fresh samples versus 135 ± 4 mg/dl and 10 ± 1.5 mg/dl in the frozen samples (P = 0.2 and 0.1, respectively). We also tested the effects of plasma filtration on laboratory controls by measuring the total whole plasma apoA-I concentration, in both filtered and unfiltered plasma samples, and found no significant difference (mean of 124 ± 21 mg/dl for the unfiltered vs.114 ± 10 mg/dl for the filtered, P = 0.1), concluding that any precipitation of lipoproteins that may or may not be occurring with plasma thawing is not removed by filtration. In addition, we tested whether any apoC-III is lost through the 10,000 Da MWCO filter as a result of apolipoprotein dissociation during the immunoaffinity chromatography steps. We separated plasma controls by anti-apoA-I columns and loaded the apoA-I-containing fraction onto anti-apoC-III columns. The eluted apoA-I containing apoC-III was then concentrated with 10,000 MWCO concentrators. The filtrates that would normally be discarded after this step were analyzed with a highly sensitive ELISA. Measurement of apoC-III was undetectable. Therefore, we conclude that apoC-III remains with the HDL particle during the preparation steps. Finally, this experiment shows that even a very small intact HDL, having a single apoA-I at 28,800 Da with some lipid and apoC-III, would not be lost through the 10,000 Da MWCO filter. Because it has been shown that apoA-I is present in LDL, we measured the apoB concentration in the elution obtained from anti-apoA-I immunoaffinity chromatography of plasma samples from laboratory controls with a high-sensitivity sandwich ELISA using affinity-purified antibodies (Academy Bio-Medical Co.) and a horseradish peroxidase/ortho-phenylenediamine detection system. This ELISA has a lower limit of detection of 0.0015 mg/dl. We found that only 2.7% of whole plasma apoB was found in the apoA-I bound fraction. More so, after further separation of apoA-I-containing lipoproteins using anti-apoE and anti-apo-C-III columns, 94% of the apoB measured did not contain apoE or apoC-III. Overall, <0.03 mg/dl of apoB associated with apoA-I had apoE, apoC-III, or both; thus the amount of apoE or apoC-III contributed by apoB lipoproteins to our apoE and apoC-III measurements is negligible. Each of the four immunofractions was then separated on a gradient gel and classified into four distinct sizes based on the nomenclature proposed by Rosenson et al. (40Rosenson R.S. Brewer Jr, H.B. Chapman M.J. Fazio S. Hussain M.M. Kontush A. Krauss R.M. Otvos J.D. Remaley A.T. Schaefer E.J. HDL measures, particle heterogeneity, proposed nomenclature, and relation to atherosclerotic cardiovascular events.Clin. Chem. 2011; 57: 392-410Crossref PubMed Scopus (360) Google Scholar) (Fig. 2). First, the eluted fractions were concentrated to a volume of 50 μl using
Basal cell carcinoma is a common skin tumor. Cold atmospheric plasma (CAP) has attracted increasing attention for its antitumor effects. The aim of the present study was to investigate the effects and related mechanisms of two CAP‑activated solutions on the TE354T basal cell carcinoma and HaCat keratinocyte cell lines. Plasma‑activated solution (PAS) was prepared by CAP irradiation of DMEM and PBS. TE354T cells were treated with PAS in vitro and the effect on cell viability was evaluated by an MTT assay. The apoptosis rate was detected by Annexin V/propidium iodide staining. Furthermore, western blotting and RNA‑sequencing were performed. The present results demonstrated that PAS induced apoptotic signaling in basal cell carcinoma cells, and that this effect was associated with the activation of the MAPK signaling pathway. Therefore, the present study demonstrated that PAS may serve as a novel treatment for basal cell carcinoma.
This work aimed to clarify the potential regulating effects of Qufeng Xuanfei formula (QFXF) on airway neurogenic inflammation and its underlying target signal pathway. Guinea pig model of airway hyperergy (AHR) was used. The relative susceptibility of major proteins to airway neurogenic inflammation was assessed using Western blot immunoassay followed by being separated by SDS-PAGE. Compared to the model group, QFXF of all concentrations effectively depressed the capsaicin enhanced cough in guinea pigs and the peak values of airway resistance significantly decreased. The results illustrated that QFXF alleviated cough symptom in guinea pigs and reduced airway neurogenic inflammation when compared to AHR model group. Airway inflammation and damage, as well as the levels of NGF, SP and c-Fos in QFXF decreased the most in the high-dose group. The mechanism of antitussive activity may be associated with reducing airway inflammation. QFXF displayed effect on chronic cough through reducing the levels of neuropeptides, attenuating airway inflammation and promoting recovery from disease to decrease the airway neuro sensitivity, suggesting that the potential mechanism may be related to Ras/ERK/c-Fos pathway.
Bone is one of the most important organs in the human body. It provides structure, function, and protection for other vital organs; therefore, bone maintenance and homeostasis are critical processes. As humans age, their bone mineral density decreases, which leads to diseases like osteoporosis. This disease affects one in two women and one in five men aged 50 and over. As the aging population increases, the interest and significance of studying this debilitating bone disease becomes more relevant. Current therapeutic products for osteoporosis have many side effects and can be taken for a limited number of years. Most therapeutic products only focus on decreasing bone resorption, not increasing bone formation. Bone morphogenetic protein 2 is an essential growth factor that drives osteoblast differentiation and activity and is essential for bone formation. However, usage in the clinic is unsuccessful due to several side effects. Recently, a signaling disparity in bone marrow stromal cells within the bone morphogenetic protein pathway that led to decreased bone morphogenetic protein 2 responsiveness was identified in patients diagnosed with osteoporosis. However, it is unclear how other cell populations, especially osteoblasts, which are key players in bone remodeling, are affected and whether the bone morphogenetic protein pathway is affected during osteoporosis. Our research group designed a novel peptide, casein kinase 2.3, that acts downstream of the bone morphogenetic receptor type Ia and increases bone mineralization in murine cells and primary bovine osteoblasts. The aim of the study presented here was to compare the responsiveness of osteoblasts to bone morphogenetic protein 2 and casein kinase 2.3, especially in patients diagnosed with osteoporosis. Mature osteoblasts were extracted from patients diagnosed with osteoporosis or osteoarthritis from Christiana Care Hospital in Newark, Delaware. They were stimulated with either bone morphogenetic protein 2 or casein kinase 2.3, and their effect on osteoblast activity was determined. The osteoporotic patients showed no mineralization response to bone morphogenetic protein 2 stimulation, while the osteoarthritis patients significantly responded to bone morphogenetic protein 2 stimulation. Furthermore, markers for osteoblast activity were increased by casein kinase 2.3, which was in sharp contrast to bone morphogenetic protein 2. This further supports a major bone morphogenetic protein signaling disparity in both the elderly and those suffering with osteoporosis. Both patient types did significantly respond to casein kinase 2.3. Further analysis of the bone morphogenetic protein pathway could lead to new therapeutic products for osteoporosis.
To study basic thyroid stimulating hormone (bTSH) levels impact on outcomes of in vitro fertilization (IVF) or intracytoplasmic sperm injection (ICSI) in Qinghai.Totally 282 cases with IVF cycles and 93 cases with ICSI cycles were studied prospectively, according to bTSH level, patients were divided into four groups. Reproduction rate, clinical pregnancy rate, miscarriage rate and live birth rate were studied among four groups.(1) In 375 cases with IVF/ICSI cycles, bTSH was positively correlated with abortion rate (r = 0.42, P = 0.04), but live birth rate and growing rate showed negative correlations with bTSH (r = -0.42, -0.28; P = 0.04, 0.03). bTSH and the number of eggs, the number of fertilized eggs, the number of embryos, biochemical pregnancy rate, and clinical pregnancy rate were no significant correlation (all P > 0.05). (2) Among women at group of ≤ 1.7, >1.7 and ≤ 2.5, >2.5 and ≤ 3.5, >3.5 mU/L, the implantation rates were 28.7%, 27.3%, 37.7% and 19.2%, live birth rates were 80.9%, 75.0%, 82.7%, and 59.8%, abortion rates were 19.0%, 15.0%, 16.7%, 40.1%; they all showed significant difference (all P < 0.05). Abortion rate in women with high bTSH level was higher than that of women with lower bTSH level, however implantation rate, live birth rate in women with high bTSH level were lower.When bTSH level is >3.5 mU/L, the abortion rate were increased, but live birth rate, rate of implantation were decreased.
The central nervous system normally functions at O2 levels which would be regarded as hypoxic by most other tissues. However, most in vitro studies of neurons and astrocytes are conducted under hyperoxic conditions without consideration of O2-dependent cellular adaptation. We analyzed the reactivity of astrocytes to 1, 4 and 9% O2 tensions compared to the cell culture standard of 20% O2, to investigate their ability to sense and translate this O2 information to transcriptional activity. Variance of ambient O2 tension for rat astrocytes resulted in profound changes in ribosomal activity, cytoskeletal and energy-regulatory mechanisms and cytokine-related signaling. Clustering of transcriptional regulation patterns revealed four distinct response pattern groups that directionally pivoted around the 4% O2 tension, or demonstrated coherent ascending/decreasing gene expression patterns in response to diverse oxygen tensions. Immune response and cell cycle/cancer-related signaling pathway transcriptomic subsets were significantly activated with increasing hypoxia, whilst hemostatic and cardiovascular signaling mechanisms were attenuated with increasing hypoxia. Our data indicate that variant O2 tensions induce specific and physiologically-focused transcript regulation patterns that may underpin important physiological mechanisms that connect higher neurological activity to astrocytic function and ambient oxygen environments. These strongly defined patterns demonstrate a strong bias for physiological transcript programs to pivot around the 4% O2 tension, while uni-modal programs that do not, appear more related to pathological actions. The functional interaction of these transcriptional 'programs' may serve to regulate the dynamic vascular responsivity of the central nervous system during periods of stress or heightened activity.
Gamma-aminobutyric acid (GABA), a non-protein-producing amino acid, is extensively found in microorganisms, plants and vertebrates, and is abundantly expressed in the spinal cord and brain. To date, GABA is considered to be the major inhibitory neurotransmitter in the central nervous system. Its physiological effects are related to the regulation of synaptic transmission, the promotion of neuronal development and relaxation, and the prevention of insomnia and depres-sion. Mediated through its specific receptors, it plays a pivotal role in the control of neuronal ex-citability, which can serve as anovel target for developing analgesics for pain management. This review provides an update on the accumulating evidence of specific GABA receptors and their subtypes in the involvement of pain analgesia.
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