OBJECTIVETo study the changes in expression and activity of protein phosphatases type 2A (PP2A ) during differentiation of NB4 and NB4-MR2 cells induced by all-trans retinoic acid (ATRA), and evaluate the role of PP2A in MR2 resistance to ATRA.

METHODSATRA, okadaic acid (OKA) and ATRA + OKA at the same dosage were incubated with NB4 and MR2 cells respectively. Wright's staining and NBT reduction test were employed to evaluate the change in the cells. The CD11b expression was measured by flow cytometry. The activity of PP2A was evaluated by serine/threonine phosphatase assay system, and the level of PP2A subunits was detected by Western blot.

RESULTS1) Wright's staining, NBT reduction test and flow cytometry results showed OKA could augment the differentiation of NB4 induced by ATRA, and OKA + ATRA induced slight differentiation of MR2 cells. 2) Phosphatase assay showed a decrease in PP2A phosphatase activity [(534 +/- 43) pmol x min(-1) x microg protein(-1)] in NB4 after ATRA treatment, accompanied with that activity [(959 +/- 83) pmol x min(-1) x microg protein(-1)] in untreated NB4 cells. OKA enhanced the inhibitory effect of ATRA on the activity in NB4. When OKA + ATRA was incubated with MR2, PP2A in the cells was significantly decreased [(229 +/- 23) pmol x min(-1) x microg protein(-1)]. 3) Western blot analysis showed that the level of PP2A catalytic subunit (PP2A/C) was decreased during the course of ATRA-induced NB4 cell differentiation, whereas expressions of every subunits of PP2A in MR2 cells were somewhat unaltered.

CONCLUSIONExpression of PP2A/C and activity of PP2A is decreased during differentiation of NB4 induced by ATRA, and no repression of the PP2 activity maybe related to MR2 resistance to ATRA.

Cell Differentiation ; drug effects ; Cell Line, Tumor ; Humans ; Leukemia, Promyelocytic, Acute ; metabolism ; pathology ; Okadaic Acid ; pharmacology ; Phosphoprotein Phosphatases ; metabolism ; Protein Phosphatase 2 ; antagonists & inhibitors ; metabolism ; Tretinoin ; pharmacology

Accidents, Traffic ; statistics & numerical data ; China ; Humans ; Wounds and Injuries ; epidemiology

BACKGROUNDFolic acid is very important for embryonic development and dihydrofolate reductase is one of the key enzymes in the process of folic acid performing its biological function. Therefore, the dysfunction of dihydrofolate reductase can inhibit the function of folic acid and finally cause the developmental malformations. In this study, we observed the abnormal cardiac phenotypes in dihydrofolate reductase (DHFR) gene knock-down zebrafish embryos, investigated the effect of DHFR on the expression of heart and neural crest derivatives expressed transcript 2 (HAND2) and explored the possible mechanism of DHFR knock-down inducing zebrafish cardiac malformations.

METHODSMorpholino oligonucleotides were microinjected into fertilized eggs to knock down the functions of DHFR or HAND2. Full length of HAND2 mRNA which was transcribed in vitro was microinjected into fertilized eggs to overexpress HAND2. The cardiac morphologies, the heart rates and the ventricular shortening fraction were observed and recorded under the microscope at 48 hours post fertilization. Whole-mount in situ hybridization and real-time PCR were performed to detect HAND2 expression.

RESULTSDHFR or HAND2 knock-down caused the cardiac malformation in zebrafish. The expression of HAND2 was obviously reduced in DHFR knock-down embryos (P < 0.05). Microinjecting HAND2 mRNA into fertilized eggs can induce HAND2 overexpression. HAND2 overexpression rescued the cardiac malformation phenotypes of DHFR knock-down embryos.

CONCLUSIONSDHFR plays a crucial role in cardiac development. The down-regulation of HAND2 caused by DHFR knock-down is the possible mechanism of DHFR knock-down inducing the cardiac malformation.

Animals ; Basic Helix-Loop-Helix Transcription Factors ; genetics ; physiology ; Female ; Heart ; embryology ; Heart Defects, Congenital ; etiology ; Tetrahydrofolate Dehydrogenase ; genetics ; physiology ; Zebrafish ; Zebrafish Proteins ; genetics ; physiology

OBJECTIVETo explore the safety and feasibility of cervical triangulating stapled anastomosis(TSA) for esophagogastric anastomosis(EGA) after minimally invasive esophagectomy (MIE).

METHODSClinical data of 137 patients undergoing MIE for esophageal cancer(EC) in our department from January 2013 to March 2014 using end to side circular stapled anastomosis (CSA, 60 cases) or end-to-end triangulating stapled anastomosis (TSA, 77 cases) in the neck were retrospectively analyzed. The short-term outcomes between the two groups were evaluated and compared.

RESULTSCervical anastomotic leakage occurred in 3 patients(3.9%) of TSA group, but in six(10.0%) of CSA group (P=0.152). The incidence of anastomotic stenosis was 1.3%(1/77) and 15.0%(9/60) in TSA and CSA group respectively(P=0.002). The median hospital stay and perioperative mortality were not significantly different between the two groups as well as postoperative respiratory and cardiovascular complications.

CONCLUSIONSTSA is a safely and effectively alternative method for EGA with lower incidence of postoperative gastrointestinal complications, especially in anastomotic stricture.

Anastomosis, Surgical ; methods ; Esophageal Neoplasms ; surgery ; Esophagectomy ; methods ; Humans ; Minimally Invasive Surgical Procedures ; methods ; Neck ; surgery ; Postoperative Complications ; Retrospective Studies ; Treatment Outcome

BACKGROUNDPrevious studies have shown that brain functional activity in the resting state is impaired in Alzheimer's disease (AD) patients. However, alterations in intrinsic brain activity patterns in mild cognitive impairment (MCI) patients are poorly understood. This study aimed to explore the differences in regional intrinsic activities throughout the whole brain between aMCI patients and controls.

METHODSIn the present study, resting-state functional magnetic resonance imaging (fMRI) was performed on 18 amnestic MCI (aMCI) patients, 18 mild AD patients and 20 healthy elderly subjects. And amplitude of low-frequency fluctuation (ALFF) method was used.

RESULTSCompared with healthy elderly subjects, aMCI patients showed decreased ALFF in the right hippocampus and parahippocampal cortex, left lateral temporal cortex, and right ventral medial prefrontal cortex (vMPFC) and increased ALFF in the left temporal-parietal junction (TPJ) and inferior parietal lobule (IPL). Mild AD patients showed decreased ALFF in the left TPJ, posterior IPL (pIPL), and dorsolateral prefrontal cortex compared with aMCI patients. Mild AD patients also had decreased ALFF in the right posterior cingulate cortex, right vMPFC and bilateral dorsal MPFC (dMPFC) compared with healthy elderly subjects.

CONCLUSIONSDecreased intrinsic activities in brain regions closely related to episodic memory were found in aMCI and AD patients. Increased TPJ and IPL activity may indicate compensatory mechanisms for loss of memory function in aMCI patients. These findings suggest that the fMRI based on ALFF analysis may provide a useful tool in the study of aMCI patients.

Aged ; Alzheimer Disease ; physiopathology ; Amnesia ; physiopathology ; Brain ; physiopathology ; Cognitive Dysfunction ; physiopathology ; Female ; Humans ; Magnetic Resonance Imaging ; Male

OBJECTIVETo establish a bortezomib-resistant myeloma cell line and to investigate its mechanism.

METHODSBortezomib-resistant NCI-H929 cell line (NCI-H929B) was obtained by stepwise increasing extracellular concentrations of bortezomib over a period of 8 months. The biological characteristics of NCI-H929 and NCI-H929B were observed. Proteins from NCI-H929B cell and NCI-H929 cell were extracted, run on two-dimensional gel electrophoresis. Matrix-assisted laser desorption ionization-time of flight (MALDI-TOF) and mass spectrometry (MS) were used to identify proteins. Western blot was used to further verify differential proteins.

RESULTBortezomib-resistant cell line NCI-H929B was established. NCI-H929B exhibits a 23.5-fold level of resistance to bortezomib as compared to the parental cell line NCI-H929. There were no significant differences in cellular biology of cell growth curve and cell cycle distribution between NCI-H929 and NCI-H929B cell lines.Whole proteins of NCI-H929 and NCI-H929B myeloma cell lines were extracted by two-dimensional gel electrophoresis. Gel-image analysis revealed that there were 17 differential protein spots. A total of 14 differential protein spots were successfully identified by MALDI-TOF-MS. The result of Western blot was consistent with 2-DE.

CONCLUSIONA bortezomib-resistant human myeloma cell line NCI-H929B was successfully established. The differentially expression of proteomes may be useful for study of the bortezomib-resistant mechanisms and the molecular markers of MM.

Antineoplastic Agents ; pharmacology ; Boronic Acids ; pharmacology ; Bortezomib ; Cell Line, Tumor ; Drug Resistance, Neoplasm ; genetics ; Humans ; Multiple Myeloma ; pathology ; Myeloma Proteins ; analysis ; Proteome ; analysis ; Pyrazines ; pharmacology ; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization ; methods

OBJECTIVETo explore the significance of C4d deposition in follicular lymphoma (FL).

METHODSThe deposition of C4d was detected in samples from 133 cases of lymphoma by immunohistochemistry and FL was studied by the double stainings of CD35/C4d, CD21/C4d and Bcl-2/C4d,respectively.

RESULTSAmong the 26 FL tissues, irregular C4d deposition was seen in 19 tumor tissues. Double staining for CD35, CD21 or Bcl-2 showed the C4d deposition was around the follicular dendritic cells (FDC). There was no significant difference between the positive rate of C4d and the degree of lymphoma. No deposition was found in the diffuse areas of FL and other type lymphomas.

CONCLUSIONC4d deposition around the follicular dendritic cell in the neoplastic follicles is a specific indicator of follicular lymphoma.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Complement C4b ; immunology ; Female ; Humans ; Immunohistochemistry ; Lymphoma, Follicular ; immunology ; pathology ; Male ; Middle Aged ; Peptide Fragments ; immunology

Objective To evaluate the diagnostic advantage of the ratio of ligamentum flavum(LF) thickness to oblique canal diameter(TODR)measured on CT images in patients with lumbar canal stenosis. Methods Seventy-one patients underwent CT and MRI examinations respectively,and they were divided into two groups,the positive group and negative group,according to the presence or absence of dural sac notch caused by the LF on bilateral parasagittal MR images.Meanwhile,50 volunteers without any symptom in the lumbar region or legs were examined by CT.TODRs were measured at the L3—S1 levels of the inferior margin of the intervertebral disc on transverse CT images,respectively.The results were further analyzed with the positive findings on MR images,clinical symptoms and physical examination,so as to find out the statistical correlation between them.Results LF thickness was(3.01?0.72)mm and TODR was 0.19?0.04 in the negative group,(3.94?0.84)mm and 0.28+0.06 in the positive group,and(3.16? 0.85)mm and 0.19?0.04 in the control group.There was significant difference between positive group and negative group or control group for LF thickness(P0.24, the sensitivity,specificity and positive predictive value were 74.8%,89.6% and 73.6% respectively. Positive correlation existed between LF thickness or TODR and clinical symptom(r=0.72,0.86,P

Based on variation of Pinus massoniana families, heritablility and correlation analysis, the contents of shikimic acid and procyanidine (heritability 0.90, 0.70), dry weight of single branch (heritability 0.60) and and leaf length (heritability 0.46) were screened out as quality, yield and harvest cost traits of Folium Pini, respectively. For the different medicinal application of Folium Pini, varied methods were chosen to estimate weight and construct index equation. Weight adjustment based.on equal emphasis were used as economic weight determining method to select the best families, and the index (accuracy 0. 936 4 and heritability 0. 881 6) obtained was a little better than that obtained by equal emphasis, and much better than that by restricted index. The superior families selected with adjustment weight and equal emphasis were No. 46, 43 and 28. Partial regression were used as economic weight determining method to select the best families,and the index obtained had the highest accuracy (0.941 5) , index heritability (0. 889 9) and the genetic gain of shikimic acid content. The superior families selected with this method were No. 46, 27 and 47. No. 46 was the best families with maximal economic benefit. Our study indicated that suitable method for estimate weight and construct index equation can be applied for better accuracy of superior families selection of P. massoniana.
Breeding ; Drugs, Chinese Herbal ; analysis ; Pinus ; chemistry ; classification ; genetics ; growth & development ; Plant Leaves ; chemistry ; Plants, Medicinal ; chemistry ; classification ; genetics ; growth & development

Objective To investigate the effect of nuclear factor erythroid 2-related factor 2(Nrf2)-autophagy pathway on the incisional pain-remifentanil-induced hyperalgesia of rat model. Methods Twenty-four male Sprague-Dawley rats were randomly allocated into three groups:saline+incisional pain group(group I),remifentanil+incisional pain group(group RI)and Nrf2 agonist t-BHQ group(group tBHQ),with 8 rats in each group.In group I and RI,normal saline at 0.1 mL/(kg· min) and remifentanilat 1 μg/(kg·min) were respectively infused into caudal vein for 60 min. Rats in group t-BHQ were injected intraperitoneally with Nrf2 agonist t-BHQ(15 mg/kg),the first time at 0.5 h before remifentanil infusion,per 12 h once,4 times in a row,the rest management was the same as group RI.Brennan incision pain model rats were constructed along with the infusions in the three groups. The thermal paw withdraw latency (PWL) and mechanical paw withdraw threshold(PWT)were measured at 24 h before the infusion(T0)and at 2 h(T1),6 h(T2),24 h(T3),48 h(T4)after the infusion. Rats were sacrificed after the tests, then the L4-6segments of signal cord were removed and the expression levels of autophagy-related proteins,microtubule associated protein 1 light chain 3Ⅱ(LC3Ⅱ),Beclin 1,Nrf2 and Nrf2 downstream molecular hemeoxygenase-1 (HO-1) were detected by Western blot assay. Results The PWT and PWL values were decreased significantly with the prolongation of the processing time in the three groups. Compared with group I, PWL and PWT values were decreased at T1-4,the expression levels of LC3Ⅱand Beclin-1 were increased while Nrf2 and HO-1 were decreased at T4in group RI (P<0.05). While compared with group RI, PWL and PWT values were increased, and the expressions of Nrf2 and HO-1 were increased, LC3Ⅱ and Beclin-1 protein were upregulated in group tBHQ (P<0.05).Conclusion The activation of Nrf2-autophagy pathway can improve the incisional pain-remifentanil induced hyperalgesia.