isolation-of-precursor-b-cell-subsets-from-umbilical-cord-blood

Isolation of Precursor B-cell Subsets from Umbilical Cord Blood

Umbilical cord blood is highly enriched for hematopoietic progenitor cells at different lineage commitment stages. We have developed a protocol for ...

To investigate the complications of procedure for prolapse and hemorrhoids (PPH) and their management.

[Modified procedure for prolapse and hemorrhoids to prevent and treat postoperative complications].

Null mutation of titin-cap (TCAP) causes limb-girdle muscular dystrophy type 2G (LGMD2G). LGMD2G patients develop muscle atrophy, and lose the ability to walk by their third decade. Previous findings suggest that TCAP regulates myostatin, a key regulator of muscle growth. We tested the hypothesis that TCAP knockdown with RNA interference will lead to differential expression of genes involved in muscle proliferation and differentiation, impairing muscle cell growth. mRNA from cultured cells treated with TCAP siRNA duplex constructs was analyzed using Northern blots and real-time RT-PCR. siRNA treatment decreased TCAP mRNA expression in differentiating muscle cells. Significant (p<0.05) decreases in mRNA were observed for myogenic regulatory factors. siRNA treatment also prevented development of the normal phenotype of muscle cells. Our findings suggest that TCAP knockdown with RNA interference alters normal muscle cell differentiation.

TCAP knockdown by RNA interference inhibits myoblast differentiation in cultured skeletal muscle cells.

NULL Mutation Titin-Cap (TCAP) Ursachen Gliederg&uuml;rtel-Muskeldystrophie Typ 2G (LGMD2G). LGMD2G Patienten entwickeln Muskelschwund und verlieren die F&auml;higkeit, ihre dritte Dekade im Vorbeigehen. Bisherigen Ermittlungen zufolge TCAP Myostatin, ein wichtiger Regulator des Muskelwachstums regelt. Wir testeten die Hypothese, die TCAP Niederschlag mit RNA-Interferenz zu differenziellen Expression von Genen beteiligt, Muskel-Proliferation und Differenzierung, wird Zelle Muskelwachstum zu beeintr&auml;chtigen. mRNA von kultivierten Zellen mit TCAP SiRNA duplex Konstrukte behandelt wurde mit Nordflecken und Real-Time RT-PCR analysiert. SiRNA Behandlung verringert TCAP mRNA Ausdruck in Muskelzellen differenzieren. Signifikante (p &lt; 0,05) mRNA wurde f&uuml;r myogener regulatorische Faktoren festgestellt. SiRNA Behandlung verhindert auch Entwicklung von der normalen Ph&auml;notyp der Muskelzellen. Unsere Einsch&auml;tzungen zufolge TCAP Niederschlag mit RNA-Interferenz normale Muskel Zelldifferenzierung &auml;ndert.

TCAP Niederschlag von RNA-Interferenz hemmt Myoblast Differenzierung in kultivierten Skelettmuskel-Zellen.

Estrogen (E2) is reported to regulate skeletal muscle mass and contractile function; whether E2 exerts its effects through estrogen receptor-alpha (ERalpha) or -beta (ERbeta) is unclear. We determined the effect of ERalpha or ERbeta elimination on muscle mass and contractile function in multiple muscles of the lower limb, muscles with different locomotor tasks and proportions of fiber types I and II: soleus (Sol), plantaris (Plan), tibialis anterior (TA), and gastrocnemius (Gast) in mature female mice. To determine E2 elimination effects on muscle, we also used aromatase (Ar) knockout (KO) and wild-type (WT) mice. ERalpha and ArKO body weights were approximately 10 and 20% higher than WT. Although muscle mass tended to show a commensurate increase in both groups, only the TA was significantly larger in ERalpha (P<0.05). Ratios of muscle mass to body mass revealed significantly lower values for Gast and TA in ArKO mice (P<0.05). Tetanic tension (Po) per calculated anatomical cross-sectional area (aCSA) in ERalpha KO was lower in TA and Gast than in WT. Lower Po/aCSA in ERalpha KO Gast and TA was also supported histologically by significantly less Po/fiber areas (P<0.05). ArKO mice also had lower Po/aCSA in Gast and TA compared with WT. ERbeta KO and WT mice were comparable in all measures. Our results support the hypothesis that E2 effects on skeletal muscle are mediated in part via the ERalpha but that E2 effects may be mediated via more than one mechanism or receptor.

Estrogen receptor-alpha and -beta and aromatase knockout effects on lower limb muscle mass and contractile function in female mice.

&Ouml;strogen (E2) wird berichtet, dass die Skelettmuskulatur Massen- und kontraktilen Funktion zu regulieren; Es ist unklar, ob E2 &uuml;bt seine Wirkung durch &Ouml;strogen-Rezeptor-Alpha (ERalpha) oder -Beta (ERbeta). Wir bestimmen die Wirkung von ERalpha oder ERbeta Beseitigung auf Masse und KONTRAKTILER Muskelfunktion in mehrere Muskeln der unteren Extremit&auml;ten, Muskeln mit verschiedenen lokomotorische Aufgaben und Proportionen der Faser Typ I und II: Schollenmuskel (Sol), Plantaris (Plan), Tibialis anterior (TA) und Wadenmuskel (Gast) in Reife weibliche M&auml;use. Um zu bestimmen, E2 Eliminierung Auswirkungen auf Muskeln, verwendeten wir auch Aromatase (Ar)-Knockout (KO) und Wildtyp (WT) M&auml;use. ERalpha und ArKO K&ouml;rper Gewichte wurden ca. 10 und 20 % h&ouml;her als WT. Obwohl Muskel Masse tendenziell eine entsprechende Zunahme beider Gruppen anzeigen nur die TA in ERalpha deutlich gr&ouml;&szlig;er war (P &lt; 0,05). Verh&auml;ltnisse von Muskel Masse um K&ouml;rper Masse offenbarten deutlich niedrigere Werte f&uuml;r Gast und TA ArKO-M&auml;usen (P &lt; 0,05). Tetanischen Spannung (Po) pro berechneten anatomische Querschnittsfl&auml;che (aCSA) in ERalpha KO lag in TA und Gast als im unteren WT-Po/aCSA in ERalpha KO Gast und TA war auch histologisch durch deutlich weniger Po/Faser-Gebieten unterst&uuml;tzt (P &lt; 0,05). ArKO M&auml;use hatte auch niedrigere Po/aCSA Gast und TA WT ERbeta KO gegen&uuml;ber und WT M&auml;use waren in alle Ma&szlig;nahmen vergleichbar. Unsere Ergebnisse st&uuml;tzen die Hypothese, die E2-Auswirkungen auf die Skelettmuskulatur werden teilweise &uuml;ber die ERalpha vermittelt, sondern die E2-Effekte k&ouml;nnen &uuml;ber mehr als ein Mechanismus oder Rezeptor vermittelt werden.

&Ouml;strogen Rezeptor-Alpha und Beta und Aromatase k.o. Auswirkungen auf die unteren Extremit&auml;ten Massen- und KONTRAKTILER Muskelfunktion bei weiblichen M&auml;usen.

The aim of this study was to investigate the analgesic effects of intravenous flurbiprofen axetil for the refractory pain in cancer patients.

Intravenous flurbiprofen axetil can increase analgesic effect in refractory cancer pain.

A novel, easy to perform PCR-based method employing specific DNA methylation biomarkers to detect B-cell neoplasms in a variety of B-cell lines and B lymphoblastic leukemia (B-ALL) patient specimens has been developed. This method detects as few as 5 B-ALL cells, or 1 B-ALL cell in 1,000,000 normal background blood cells using a single marker, DLC-1 gene CpG island (CGI) methylation. By adding two additional markers PCDHGA12 and RPIB9, over 80% of B-ALL cases were detected in patients' bone marrow and/or peripheral blood specimens. We have traced clinical B-ALL cases up to 10 years retrospectively and the DLC-1 methylation is correlated with patient clinical status. Thus, this epigenetic-based molecular method demonstrates its potential use in the diagnosis of B-cell neoplasia, in addition to traditional approach such as clinical features, morphology, immunophenotype, and genetic analysis.

Molecular detection of B-cell neoplasms by specific DNA methylation biomarkers.

In contrast to experimental evidence regarding the postorbital bar, postorbital septum, and browridge, there is exceedingly little evidence regarding the load-bearing nature of soft-tissue structures of the mammalian circumorbital region. This hinders our understanding of pronounced transformations during primate origins, in which euprimates evolved a postorbital bar from an ancestor with the primitive mammalian condition where only soft tissues spanned the lateral orbital margin between frontal bone and zygomatic arch. To address this significant gap, we investigated the postorbital microanatomy of rabbits subjected to long-term variation in diet-induced masticatory stresses. Rabbits exhibit a masticatory complex and feeding behaviors similar to primates, yet retain a more primitive mammalian circumorbital region. Three cohorts were obtained as weanlings and raised on different diets until adult. Following euthanasia, postorbital soft tissues were dissected away, fixed, and decalcified. These soft tissues were divided into inferior, intermediate, and superior units and then dehydrated, embedded, and sectioned. H&E staining was used to characterize overall architecture. Collagen orientation and complexity were evaluated via picrosirius-red staining. Safranin-O identified proteoglycan content with additional immunostaining performed to assess Type-II collagen expression. Surprisingly, the ligament along the lateral orbital wall was composed of elastic fibrocartilage. A more degraded organization of collagen fibers in this postorbital fibrocartilage is correlated with increased masticatory forces due to a more fracture-resistant diet. Furthermore, the lack of marked changes in the extracellular composition of the lateral orbital wall related to tissue viscoelasticity suggests it is unlikely that long-term exposure to elevated masticatory stresses underlies the development of a bony postorbital bar.

Masticatory loading, function, and plasticity: a microanatomical analysis of mammalian circumorbital soft-tissue structures.

The adaptive growth response of cartilage, or chondral modeling, can result in changes in joint and limb proportions during ontogeny and ultimately contribute to the adult form. Despite Hamrick's (1999) reevaluation of the mechanisms of chondral modeling, the process of chondral modeling remains poorly studied in animal models. Here, we characterize the macro- and microanatomical responses of the femoral growth plate, articular cartilage, and bone in 15 juvenile Sus scrofa domestica subjected to different locomotor activity patterns. The exercised animals exhibit thinner cartilage zones, greater cellularity and larger proliferative chondrocyte areas in the growth plate, as well as larger femoral dimensions and a more elongate femoral head compared with sedentary controls. In general, the growth plate demonstrates greater adaptive changes than articular cartilage. Moreover, chondrocyte hypertrophy and proliferation were found to be responsive to locomotor loading and thus more important factors in chondral modeling than the extracellular matrix variables that were examined herein. In sum, the underlying mechanisms of adaptive chondrogenesis and bone plasticity are key to informing evolutionary and translational studies regarding determinants of variation in joint form and function. Given the disparity between the predictions of chondral modeling theory and our experimental findings, this suggests a need for further evaluation of chondral modeling responses during ontogeny.

Mammalian limb loading and chondral modeling during ontogeny.

Bisphosphonates (BPs) like Zometa (ZA) are widely used to treat complications of bony metastases in cancer patients. A serious adverse event occurs in 1-12% of patients on BP therapy, osteonecrosis of the jaw (BPONJ). BPONJ develops after oral trauma and is manifested by poor wound healing and soft-tissue breakdown followed by exposure and necrosis of intra-oral bone. Currently, there is no effective clinical treatment for BPONJ.

Bisphosphonate effects on the behaviour of oral epithelial cells and oral fibroblasts.

It has previously been known that transcription of the PGC-1Î± gene can be either inhibited or stimulated by p38 MAP kinase (p38 MAPK). To determine whether p38 MAPK plays an inhibitory or stimulatory role in PGC-1Î± gene transcription, we further investigated the role of p38 MAPK in this study. Our results showed that the basal level of p38 MAPK phosphorylation was increased in gastrocnemius of mice under HFD and that p38 MAPK stimulated PGC-1Î± gene transcription in C(2)C(12) myotubes. Our results also provided new mechanisms in myotubes that the p38 MAPK-induced PGC-1Î± gene transcription was mediated by CREB. In exploring the role of the Akt-dependent insulin signaling on PGC-1Î± gene transcription, we found that the basal Akt-dependent signaling was increased in gastrocnemius of mice under HFD. The p38 MAPK-induced PGC-1Î± gene transcription was prevented by insulin. Insulin suppression of PGC-1Î± gene transcription was neutralized by overexpression of the constitutively nuclear form of FoxO1. Finally, we located three insulin response elements (IREs) in the PGC-1Î± promoter, and mutations of these IREs abolish or blunt activity of the PGC-1Î± promoter. Together, our results show that transcription of the PGC-1Î± gene is balanced by different intracellular signaling pathways.

Fine-tuned regulation of the PGC-1Î± gene transcription by different intracellular signaling pathways.

The main goal of this review is to provide more specific and effective targets for prevention and treatment of insulin resistance and associated atherosclerosis. Modern technologies and medicine have vastly improved human health and prolonged the average life span of humans primarily by eliminating various premature deaths and infectious diseases. The modern technologies have also provided us abundant food and convenient transportation tools such as cars. As a result, more people are becoming overfed and sedentary. People are generally ingesting more calories than their bodies' need, leading to the so-called "positive energy imbalance", which is inseparable from the development of insulin resistance and its associated atherosclerosis. A direct consequence of insulin resistance is hyperinsulinemia. The current general view is that insulin is not functional properly in the presence of insulin resistance. Thus, the role of insulin itself in the development of insulin resistance and associated atherosclerosis has not been recognized. We have recently observed that the basal level of insulin signaling is increased in the presence of insulin resistance and hyperinsulinemia. In this review, we will explain how the increased basal insulin signaling contributes to the development of insulin resistance and associated atherosclerosis. We will first explain how insulin causes insulin resistance through two arbitrary stages (before and after the presence of obvious insulin resistance), and, then, explain how the excess exposure to insulin and the relative insulin insufficiency contributes to the atherosclerotic diseases. We propose that blockade of the excess insulin signaling is a viable approach to prevent and/or reverse insulin resistance and its associated atherosclerosis.

Excess exposure to insulin is the primary cause of insulin resistance and its associated atherosclerosis.

The role of the IRE1Î±-X-box-binding protein 1 (XBP1) pathway in the insulin-mediated hepatic lipogenic program and associated mechanisms were investigated in this study. We observed that phosphorylation of IRE1Î± (an upstream activator of XBP1) and splicing (activation) of XBP1 were elevated in the liver of the C57BL/6 mice with insulin resistance/hyperinsulinemia induced by high-fat diet. Treatment of nonobese diabetic mice with insulin activated hepatic XBP1. In cultured primary mouse hepatocytes, prolonged exposure to insulin induced IRE1Î± phosphorylation and XBP1 splicing significantly in the presence of insulin resistance. Overexpression of the activated XBP1 elevated the promoter activities of the sterol regulatory element-binding protein (SREBP)-1c and fatty acid synthase (FAS) genes. Knockdown of either the IRE1Î± or XBP1 gene by small interfering RNA prevented the insulin-stimulated promoter activities of both SREBP-1 and FAS genes. In investigating the associated mechanisms, we found a direct interaction between XBP1 and SREBP-1 promoter detected by the chromatin immunoprecipitation assays. Furthermore, the XBP1-mediated stimulation of the FAS promoter was eliminated by knocking down the SREBP-1c gene (Srebf1). Finally, we observed that insulin activation of the IRE1Î±-XBP1 pathway was prevented by inhibition of mammalian target of rapamycin-dependent protein synthesis. In conclusion, our results show that the IRE1Î±-XBP1-mediated unfolded protein response pathway is an integrated part of the insulin-induced hepatic lipogenic program and functions at an increased basal level in the presence of insulin resistance and hyperinsulinemia. Besides, the insulin-mediated protein synthesis is tightly connected with the insulin-mediated lipogenic program.

Constitutive role for IRE1Î±-XBP1 signaling pathway in the insulin-mediated hepatic lipogenic program.

The primary player that induces insulin resistance has not been established. Here, we studied whether or not fat can cause insulin resistance in the presence of insulin deficiency. Our results showed that high-fat diet (HFD) induced insulin resistance in C57BL/6 (B6) mice. The HFD-induced insulin resistance was prevented largely by the streptozotocin (STZ)-induced moderate insulin deficiency. The STZ-induced insulin deficiency prevented the HFD-induced ectopic fat accumulation and oxidative stress in liver and gastrocnemius. The STZ-induced insulin deficiency prevented the HFD- or insulin-induced increase in hepatic expression of long-chain acyl-CoA synthetases (ACSL), which are necessary for fatty acid activation. HFD increased mitochondrial contents of long-chain acyl-CoAs, whereas it decreased mitochondrial ADP/ATP ratio, and these HFD-induced changes were prevented by the STZ-induced insulin deficiency. In cultured hepatocytes, we observed that expressions of ACSL1 and -5 were stimulated by insulin signaling. Results in cultured cells also showed that blunting insulin signaling by the PI3K inhibitor LY-294002 prevented fat accumulation, oxidative stress, and insulin resistance induced by the prolonged exposure to either insulin or oleate plus sera that normally contain insulin. Finally, knockdown of the insulin receptor prevented the oxidative stress and insulin resistance induced by the prolonged exposure to insulin or oleate plus sera. Together, our results show that insulin and insulin signaling are required for fat induction of insulin resistance in mice and cultured mouse hepatocytes.

Article	Year
[Modified procedure for prolapse and hemorrhoids to prevent and treat postoperative complications]. Di 1 jun yi da xue xue bao = Academic journal of the first medical college of PLA\| PubMed ID: 16109572	2005
TCAP knockdown by RNA interference inhibits myoblast differentiation in cultured skeletal muscle cells. Neuromuscular disorders : NMD\| PubMed ID: 18440815	2008
Estrogen receptor-alpha and -beta and aromatase knockout effects on lower limb muscle mass and contractile function in female mice. American journal of physiology. Endocrinology and metabolism\| PubMed ID: 19176355	2009
Intravenous flurbiprofen axetil can increase analgesic effect in refractory cancer pain. Journal of experimental & clinical cancer research : CR\| PubMed ID: 19267934	2009
Molecular detection of B-cell neoplasms by specific DNA methylation biomarkers. International journal of clinical and experimental pathology\| PubMed ID: 20224725	2010
Masticatory loading, function, and plasticity: a microanatomical analysis of mammalian circumorbital soft-tissue structures. Anatomical record (Hoboken, N.J. : 2007)\| PubMed ID: 20235321	2010
Mammalian limb loading and chondral modeling during ontogeny. Anatomical record (Hoboken, N.J. : 2007)\| PubMed ID: 20235322	2010
Bisphosphonate effects on the behaviour of oral epithelial cells and oral fibroblasts. Archives of oral biology\| PubMed ID: 21146154	2011
Fine-tuned regulation of the PGC-1Î± gene transcription by different intracellular signaling pathways. American journal of physiology. Endocrinology and metabolism\| PubMed ID: 21156859	2011
Excess exposure to insulin is the primary cause of insulin resistance and its associated atherosclerosis. Current molecular pharmacology\| PubMed ID: 21241239	2011
Constitutive role for IRE1Î±-XBP1 signaling pathway in the insulin-mediated hepatic lipogenic program. Endocrinology\| PubMed ID: 21447637	2011
Insulin and insulin signaling play a critical role in fat induction of insulin resistance in mouse. American journal of physiology. Endocrinology and metabolism\| PubMed ID: 21586696	2011

Jie Ning

Biography

JoVE Journal Publications

Other Publications