Objective To explore the effect of sodium butyrate(NaB) on phosphorylation/ acetylation of histone H3(ph/acH3) at G?-globin gene and A?-globin gene promoter regions in K562 cells.Methods K562 cells were devided into 2 groups:K562 cells were grown in the presence or absence of 0.5 mmol?L-1NaB for 48 h [K562(NaB) group] and untreated K562 cells group(K562 group).Semi-quantitative RT-PCR was employed to measure the levels of G?-globin mRNA and A?-globin mRNA.The real time PCR-based chromatin immunoprecipitation(ChIP) was used to detect the levels of ph/acH3 at G?-globin gene and A?-globin gene promoter regions.Results Compared with the K562 group,there was a 1.4-fold(t=-149.022,P=0.000) and 1.2-fold(t=-13.363,P=0.000) increase in G?-globin mRNA and A?-globin mRNA,respectively,in K562(NaB) group.The level of ph/acH3 at G?-globin gene and A?-globin gene promoter region increased by 2.9-fold(t=-12.833,P=0.006) and 3.2-fold(t=-10.484,P=0.000),respectively,in K562(NaB) group,compared with the K562 group.The %Input value of G?-globin and A?-globin promoter fragment was 10.0-fold(P=0.000) and 9.5-fold(P=0.000) higher than that value of Necdin gene promoter fragment in the K562(NaB) group,while the %Input value of G?-globin and A?-globin promoter fragment was 3.2-fold(P=0.000) and 2.7-fold(P=0.000) higher than that value of necdin gene promoter fragment in K562 group.Conclusions NaB improves the phosphorylation and acetylation of H3 at ?-globin gene promoter regions,and this may be one of the mechanisms of expression of ?-globin genes induced by NaB.

OBJECTIVETo develop a real-time PCR-based chromatin immunoprecipitation (ChIP) assay for determining the effect of sodium butyrate on acetylation of histone in gamma-globin gene promoter regions in K562 cells.

METHODSK562 cells were grown in the presence or absence of 0.5 mmol/L sodium butyrate for 48 h, and 1=10(7) cells per group were used for real-time PCR-based ChIP with anti-acetylated histone H3 or H4 antibodies. The levels of acetylated histone H3 and H4 (acH3 and acH4) in Ggamma- and Agamma-globin gene promoter regions were measured.

RESULTSIn the K562 cells with sodium butyrate treatment or without any treatment, the levels of acH3 or acH4 in Ggamma- or Agamma-globin gene promoter were higher than that in the necdin gene (negative control). Compared with the untreated K562 cells, the cells treated with 0.5 mmol/L sodium butyrate showed a 3.1-fold or 2.6-fold increase in acH3 or acH4 in Ggamma-globin gene promoter region, with also a 3.7-fold or 3.2-fold increase in acH3 or acH4 in Agamma-globin gene promoter region, respectively (P<0.01).

CONCLUSIONWe have successfully developed a real-time PCR-based ChIP assay for analyzing the acetylation of histone H3 and H4 in gamma-globin gene promoter regions. Our results support the role of sodium butyrate in increasing the level of acetylated histone in gamma-globin gene promoter regions.

Acetylation ; Butyrates ; pharmacology ; Chromatin Immunoprecipitation ; methods ; Histones ; chemistry ; Humans ; K562 Cells ; Promoter Regions, Genetic ; genetics ; Real-Time Polymerase Chain Reaction ; methods ; gamma-Globins ; genetics

Objective To analyze the patterns of cortical atrophy of the two subtypes of frontotemporal lobar degeneration (FTLD ),behavioural-vsriant frontotemporal dementia (bvFTD ) and primary progressive aphasia (PPA).And to compare them with that of Alzheimer disease (AD) to provide an objective basis for early diagnosis and differential diagnosis.MethodsA total of 83 patients were enrolled in this study and there were 30 patients with cognitively normal controls (CN),30 with AD and 23 with FTLD (10 with bvFTD,13 with PPA).Philips 3.0 T TX scanner and 8 channel head coil was employed.Three dimensional turbo fast echo(3D-TFE)T1WI sequence with high resolution was used to collect the volume data of gray matter.3D-TFE T1 WI images were normalized and segmented into gray matter map for statistical analysis by SPM 8 and VBM 8.The false discovery rate (FDR) was adopted in P value adjustment,P ＜ 0.001,and the cluster size was set at 5.The full width at half maximum (FWHM ) was set at 4 mm for the smoothing.Paired t test was used for statistics.ResultsIn bvFTD,PPA and AD groups,there were diffuse regions with reduced volume in cerebral cortex and subcortical structures (such as the hippocampus,the amygdala,the caudate nuclei,et al).The most obvious atrophic region in bvFTD and PPA group was found in the frontotemporal.Compared with AD,gray matter atrophy in bvFTD was found in brain regions including bilateral temporal lobes,bilateral superior temporal pole gyri,bilateral middle temporal pole gyri,right fusiform gyrus and bilateral frontal lobes.Among them,temporal and frontal lobes atrophy had obvious right partial lateralizing,with 14 301 voxels in right temporal lobe and 5105 in left (t =-5.03,P＜0.05).The number of atrophy voxels in right and left frontal lobe were 1344 and 125 (t =3.45,P ＜0.05).The left temporooccipital lobe atrophy was more obvious than the right in PPA,with 15 637 voxels in left and 10 723 in right ( t =- 2.65,P ＜ 0.05 ).ConclusionsThere are different brain gray matter atrophy patterns in bvFD,PPA and AD.Among them,bvFTD has asymmetric right frontal and temporal lobe atrophy,which may be related to characteristic personality changes.On the other hand,the asymmetric atrophy in left temporooccipital lobe may be responsible for the aphasis of patients with PPA.

BACKGROUNDMu opioid receptor plays an important role in many physiological functions. Fentanyl is a widely used opioid receptor agonist for analgesia. This study was conducted to test the role of mu-opioid receptor on insulin release by determining whether fentanyl affected insulin release from freshly isolated rat pancreatic islets and if small interfering RNAs (siRNA) targeting mu-opioid receptor in the islets could knock down mu-opioid receptor expression.

METHODSIslets were isolated from ripe SD rats' pancreas by common bile duct intraductal collagenase V digestion and purified by discontinuous Ficoll density gradient centrifugation. The siRNA knock-down of mu-opioid receptor mRNA and protein in islet cells was analyzed by semi-quantitative real time-PCR and Western blotting. After siRNA-transfection for 48 hours, the islets were co-cultured with fentanyl as follows: 0 ng/ml, 3 ng/ml and 30 ng/ml for 48 hours. Then glucose-evoked insulin release was performed. As a control, the insulin release was also analyzed in islets without siRNA-trasfection after being co-cultured with fentanyl for 48 hours.

RESULTSAfter 48 hours of transfections, specific siRNA targeting of mu-opioid receptors produced significant reduction of mu-opioid receptor mRNA and protein (P < 0.01). Fentanyl significantly inhibited glucose-evoked insulin release in islets in a concentration dependent manner (P < 0.01). But after siRNA-transfection for 48 hours, the inhibition on glucose-evoked insulin release was reversed (P < 0.01).

CONCLUSIONSRNA interference specifically reduces mu-opioid receptor mRNA and protein expression, leading to reversal of the fentanyl-induced inhibition on glucose-evoked insulin release of rat islets. The activation of opioid receptor induced by fentanyl functions to inhibit insulin release. The use of RNAi presents a promising tool for future research in diabetic mechanisms and a novel therapy for diabetes.

Analgesics, Opioid ; pharmacology ; Animals ; Cell Survival ; drug effects ; Cells, Cultured ; Fentanyl ; pharmacology ; Glucose ; pharmacology ; Insulin ; secretion ; Islets of Langerhans ; drug effects ; secretion ; Male ; RNA Interference ; RNA, Messenger ; analysis ; Rats ; Rats, Sprague-Dawley ; Receptors, Opioid, mu ; antagonists & inhibitors ; genetics ; physiology

Contracture and spasticity of ankle joints were major sources of disability in neurological impairment including stroke and cerebral palsy, etc. The manual stretching used in physical therapy might be laborious and time-consuming to the therapists and the outcome was dependent on the experience and the subjectiveend feelingof the therapists. A device was developed that could safely stretch the an-kle joint to its extreme positions with quantitative control of the resistance torque and stretching velocity. Furthermore, it could satisfy a strong need for quantitative and objective measures of the impairment and rehabilitation outcome. This was just the meaning intelligent stretching referred to. This article described the origin of the concept of intelligent stretching and its definition, operational principle, and su-periority and weakness, as well as its application in ankle joint spasticity and contracture in patients with stroke and cerebral palsy.

Objective To explore the application of erlotinib based targeting fluorescent probe in the detection of lung cancer.Methods The erlotinib based targeting fluorescent probe was prepared.The lung cancer cells of A-549 were selected as experimental group,and cervical cancer cells of CaSki,SiHa and C33-A were selected as control group.The A-549,CaSki,SiHa and C33-A cells were identified by 0.1 × 10-6,1 × 10-6,10 × 10-6 mol · L-1 fluorescent probe;the identification ability of erlotinib based targeting fluorescent probe on cells in the two groups was observed under the inverted fluorescence microscope.Results When the concentration of fluorescent probe was 10 × 10-6 mol · L-1,the strong fluorescence signal was observed in A-549 and CaSki cells,but the fluorescence signal was not observed almost in SiHa and C33-A cells.When the concentration of fluorescent probe was 1 × 10-6 mol · L-1,the strong fluorescence signal was observed in A-549 cells;the weak fluorescence signal was observed in C33A cells;the fluorescence signal was not observed almost in SiHa and C33-A cells.When the concentration of fluorescent probe was 0.1 × 10-6 mol · L-1,the strong fluorescence signal was observed in A-549 cells;the fluorescence signal was not observed almost in CaSki,SiHa and C33-A cells.Conclusion Erlotinib based targeting fluorescent probe can specifically recognize lung cancer A-549 cells.

BACKGROUNDMagnetic resonance imaging (MRI) is the most sensitive technique for evaluating the healing process and should be performed before the patients return to their exercise routines. The aim of this research was to diagnose chronic lumbago associated with lumbar muscle strain and to monitor healing process by MRI.

METHODSSixty-fve symptomatic cases of chronic lumbago caused by lumbar muscle strain were collected from March 2009 to October 2011. MRI was used to examine, diagnose and monitor the healing process. The control group included 65 random cases of asymptomatic volunteers. MRI methods included routine sequences of GRE T1WI, TSE T2WI and special sequences of T2-STIR-FS, combined with DWI. We compared the MRI characteristics of symptomatic cases before and after healing and with asymptomatic controls.

RESULTSThe important MRI characteristics of chronic lumbago with lumbar muscle strain included: (1) The low back muscle showed edema. (2) The low back intermuscular spaces showed edema and/or fluid. (3) The low back spaces beside the spinous process showed edema and/or fluid. (4) The low back vertebral articular process fossae or transverse process fossae showed fluid. Of these image characteristics, the intermuscular space edema provided the best diagnostic sensitivity, Se = 83%, with YI = 0.63, p = 74%. The low back muscle edema provided the best diagnostic specificity, Sp = 100%, with YI = 0.66, Π = 83%. And the spaces edema beside the spinous process provided the best diangnostic accuracy, Π = 86%, with YI = 0.71, Se = 80%, Sp = 91%. The diagnosis accurate could be improved by combining multiple MRI characteristics. The diagnostic accuracy could achieve Π = 93%, with YI = 0.86, Se = 100% and Sp = 86% when two characteristics were combined. After rehabilitation care, the edema disappeared on the repeated MRI.

CONCLUSIONSMRI may well be a useful diagnostic method for lumbago with lumbar muscle strain. Combining routine sequences with T2-STIR-FS and DWI sequences could demonstrate the pathological changes of lumbar muscle strain and monitor the healing.

Adult ; Chronic Disease ; Diffusion Magnetic Resonance Imaging ; Female ; Humans ; Low Back Pain ; diagnosis ; etiology ; Magnetic Resonance Imaging ; methods ; Male ; Middle Aged ; Sprains and Strains ; complications ; Wound Healing

This study investigated the relationship between serum thyrotrophin levels and dyslipidemia in subclinical hypothyroid and euthyroid subjects. A total of 110 subjects with subclinical hypothyroidism and 1,240 euthyroid subjects enrolled in this study. Patients with subclinical hypothyroidism had significantly lower high density lipoprotein cholesterol (HDL-C) levels than those who were euthyroid. The lipid profiles were each categorized and mean thyrotrophin levels were higher in subjects in the dyslipidemia subclasses than subjects in the normal subclasses. Thyrotrophin was positively associated with serum triglyceride and negatively associated with serum HDL-C in women. Thyrotrophin was also positively associated with total cholesterol (TC) in the overweight population along with TC and LDL-C in overweight women. In the euthyroid population, thyrotrophin was positively associated with TC in the overweight population. In conclusion, serum thyrotrophin was correlated with dyslipidemia in subclinical hypothyroid and euthyroid subjects; the correlation was independent of insulin sensitivity.
Adolescent ; Adult ; Aged ; Aged, 80 and over ; Cholesterol/blood ; Cholesterol, HDL/blood ; Dyslipidemias/*blood ; Female ; Humans ; Insulin Resistance ; Lipids/blood ; Male ; Middle Aged ; Thyrotropin/*blood ; Triglycerides/blood ; Young Adult

BACKGROUND:It has been reported that 70％ of patients with chronic myeloid leukemia (CML) are negative for cytogenetic and genetic markers within 1-5 months after allogeneic hematopoietic stem cell transplantation (allo-HSCT),but there are still some patients who have repeatedly varied outcomes in cytogenetic and genetic marker detection.Overall,the negative rate is up to 89.5％ at 3-12 months after allo-HSCT.OBJECTIVE:To monitor the changes in cytogenetic and genetic marker expression and to explore the prognostic significance in CML patients undergoing allo-HSCT.METHODS:Seventeen CML patients who had undergone allo-HSCT were enrolled.Chromosome G banding pattern of the bone marrow from these patients were analyzed using short-term culture method and direct method at 30 days,2,3,4,6,12,24,36,48,60,72 months after allo-HSCT.Dual-color fluorescence in situ hybridization was used to detect bcr-abl fusion gene;bcr-abl expressions in primary bone marrow ceils from CML patients were detected using RQ-PCR.Results and conclusion:There were 8/17 cases of male patient/male donor and 7/17cases of male patient/female donor (compatriots).46XX karyotype (women) was detected by multiple reexaminations after transplantation,and there was no Y chromosome or other aberration of chromosome karyotype in their karyotype.Among the 17 cases,1 case of female patient/female donor (compatriots) and 1 case of female patient/male donor (unrelated) manifested 46 XY chromosome karyotype and bcr-abl positive at 1 month after transplantation;after 4 months,these two cases still maintained 46 XY chromosome karyotype but bcr-abl negative;after 4-96 months,the karyotype continued to remain as 46 XY,and bcr-abl (-).Among the 17 cases,1 case of male patient/male donor of full-matched compatriot (brother) manifested that Ph chromosomal bcr-abl gene continuously expressed within 1-12 months after allo-HSCT;then the cases was given donor lymphocyte infusion,and the bcr-abl expression returned to be negative at 48 months after transplantation.To conclude,chromosomal karyotype analysis and bcr-abl fusion gene monitoring provide important reference value for subsequent treatment options and prognosis judgment for CML patients with allo-HSCT.

Objective To investigate the significance of the expression of phosphatidic acid phosphatase type 2 domain containing 1A(PPAPDC1A) in human colorectal cancer cell lines.Methods The high metastatic potential cells LOVO,SW620 and low metastatic potential cells SW480,RKO,HCT116 and DLD-1 were cultured,the expression of PPAPDC1A mRNA and protein in different colorectal cancer cells in logarithmic growth period was detected by real-time quantitative polymerase chain reaction and Western blot.Results There were significant differences in the expressions of PPAPDC1A mRNA and protein among the six human colorectal cancer cells (F =41.213,344.1 16;P ＜ 0.05).The expression of PPAPDC1 A mRNA and protein in highly metastatic potential cells LOVO and SW620 was significantly higher than that in DLD-1,HCT116,RKO and SW480 cells (P ＜0.05).The expression of PPAPDC1A protein in LOVO cells with high metastatic potential was significantly higher than that in SW620 cells(P ＜ 0.05).The expression of PPAPDC1A protein in DLD-1 cells was significantly higher than that in HCT116,RKO and SW480 cells (P ＜0.05).The expression of PPAPDC1 A protein in HCT116 cells with low metastatic potential was significantly higher than that in RKO and SW480 cells (P ＜ 0.05).The expression of PPAPDC1 A protein in RKO cells was significantly higher than that in SW480 cells (P ＜ 0.05).There was no significant difference in the expression of PPAPDC1A mRNA between LOVO and SW620 cells (P ＜ 0.05).There was no significant difference in the expression of PPAPDC1A mRNA between SW480,RKO,HCT116 and DLD-1 cells (P＜ 0.05).Conclusion PPAPDC1A expresses differentially in colorectal cancer cell lines,which may be involved in the invasion and metastasis of colorectal cancer.