Objective This study aimed at observing expression and clinical significance of metadherin in gastric adenocarcinoma and exploring the potentially regulating mechanism of metadherin in invation and migration of gastric cancer.Methods Expressions of metadherin and E-cadherin in primay lesion of gastric cancer were detected by immunohistochemistry and their correlation to clinicopathologic characteristics and prognosis were analyzed by Chi-square tests.Transwell assay and wound healing assay were applied for the ability of invasion and metastasis in gastric cancer cells.Then,the down-regulatied metadherin expression in MKN45 cells by RNA interference (siRNA) was carried out and furthermore,the regulation role of metadherin in epithelial-mesenchymal transition was analyzed also in invasion and migration of gastric cancer cells.Results The positive expression of metadherin was correlated to invading depth (P =0.029),lymph node metastasis (P =0.001),TNM stage (P =0.014) and inhibiting E-cadherin expression (P =0.001).The patients with positive metabherin shared poorer prognosis.Furthermore,the down-regulated metabherin in MKN45 cells would result in the increasing expression of E-cadherin,as well as decreasing expression of N-eadherin,Slug and Snail.At the same time,the abilities of invasion (P =0.027) and migration (P =0.008) of MKN45 cells was decreased.Conclusion metabherin induces EMT in metastasis of gastric adenocarcinoma via activating either Slug or Snail but not twist,which would result in the poorer prognosis.

OBJECTIVETo explore the possibilities of bone marrow stromal cells (MSCs) to adopt Schwann cell phenotype in vitro and in vivo in SD rats.

METHODSMSCs were obtained from tibia and femur bone marrow and cultured in culture flasks. Beta-mercaptoethanol followed by retinoic acid, forskolin, basic-FGF, PDGF and heregulin were added to induce differentiation of MSCs'. Schwann cell markers, p75, S-100 and GFAP were used to discriminate induced properties of MSCs' by immunofluorescent staining. PKH-67-labelled MSCs were transplanted into the mechanically injured rat sciatic nerve, and laser confocal microscopy was performed to localize the PKH67 labelled MSCs in the injured sciatic nerve two weeks after the operation. Fluorescence PKH67 attenuation rule was evaluated by flow cytometry in vitro.

RESULTSMSCs changed morphologically into cells resembling primary cultured Schwann cells after their induction in vitro. In vivo, a large number of MSCs were cumulated within the layer of epineurium around the injured nerve and expressed Schwann cell markers, p75, S-100, and GFAP.

CONCLUSIONMSCs are able to support nerve fiber regeneration and re-myelination by taking on Schwann cell function, and can be potentially used as possible substitutable cells for artificial nerve conduits to promote nerve regeneration.

Animals ; Biomarkers ; analysis ; Bone Marrow Cells ; cytology ; Cell Differentiation ; Cells, Cultured ; Flow Cytometry ; Fluorescent Antibody Technique, Indirect ; Fluorescent Dyes ; Glial Fibrillary Acidic Protein ; analysis ; Morphogenesis ; Organic Chemicals ; analysis ; Phenotype ; Rats ; Receptor, Nerve Growth Factor ; analysis ; S100 Proteins ; analysis ; Schwann Cells ; cytology ; metabolism ; Sciatic Nerve ; cytology ; injuries ; Stromal Cells ; cytology ; metabolism ; transplantation

Pineapple (Ananas comosus) leaves contain mainly phenolic components with antioxidant and hypolipidemic effects. One of the principle components is p-coumaric acid. In this study, the transport behavior of p-coumaric acid, was observed after the administration of pineapple leaf phenols in vitro. Simultaneously, the effect of the phenols on glucose, total cholesterol and triglycerides transportation and metabolism in HepG2 cells was also observed. The results showed that the phenols had good transport characteristics. 5 min after the administration, p-coumaric acid of the phenols could be detected, and the content of p-coumaric acid reached the peak concentration after 60 min of the administration. p-coumaric acid of phenols have time-and dose-dependent manner. While promoting glucose transporter (GLUT4) and low density lipoprotein receptor (LDLR) expression, the phenols decreased intracellular lipid content. This reduction of intracellular lipid content was highly correlated with the promotion of lipoprotein lipase (LPL) and hepatic triglyceride lipase (HTGL) expression, while the reduction of intracellular glucose levels was correlated with glycogen synthesis in the cells.
Ananas ; chemistry ; Biological Transport ; drug effects ; Cholesterol ; metabolism ; Glucose ; metabolism ; Hep G2 Cells ; Humans ; Lipid Metabolism ; drug effects ; Plant Extracts ; pharmacology ; Plant Leaves ; chemistry

OBJECTIVETo evaluate the effect of different CO2 pneumoperitoneum on IL-1β and IL-6 in abdominal cavity.

METHODSFifty-six female SD rats were randomly divided into seven groups. One group was served as control and the others received CO2 pneumoperitoneum. Pneumoperitoneum was established at 0.67 kPa and 1.0 L/min gas flow for 1, 2 or 3 h with CO2 (group C1 h, C2 h, and C3 h, respectively). CO2 pneumoperitoneum was further established at 1.07 kPa and 1.0 L/min gas flow for 1 h (group C8p), at 0.67 kPa and 2.0 L/min gas flow for 1 h(group C2f), and at 0.67 kPa and 3.0 L/min gas flow for 1 h (group C3f). After the procedures, peritoneal fluid was collected to analyze the IL-1β and IL-6 level by ELISA method.

RESULTSCO2 pneumoperitoneum caused peritoneal inflammatory reaction. With the increasing of duration and gas flow in CO2 pneumoperitoneum, the concentrations of IL-1β and IL-6 in group C2 h, C3 h and C3f were higher than those in group C1 h (P<0.05). On the other hand, the concentrations of IL-1β and IL-6 in peritoneal fluid did not change significantly when pressure was increased (P>0.05).

CONCLUSIONSThe inflammatory reaction in abdominal cavity after CO2 pneumoperitoneum may be attributed to duration and gas flow instead of the pressure within the standard pneumoperitoneum working pressures. Surgeons should reduce surgical duration and adopt low-velocity gas flow within normal working pressures in clinical practice.

Abdominal Cavity ; Animals ; Carbon Dioxide ; Female ; Interleukin-1beta ; metabolism ; Interleukin-6 ; metabolism ; Pneumoperitoneum, Artificial ; methods ; Rats ; Rats, Sprague-Dawley

BACKGROUNDPrevious work has shown that optic nerve and sciatic nerve conditional medium had neurotrophic activity on neurons. In order to find if the optic nerve conditioned media (CM) had a similar activity to make PC12 cells differentiate as sciatic nerve CM did, we explored the neurotrophic activity in optic nerve CM in the same in vitro system and compared the neurotrophin expression levels in optic and sciatic nerves under both conditions.

METHODSPC12 cells were used to examine the effects of neurotrophins secreted by the sciatic nerve and optic nerve. RT-PCR and real-time QPCR showed that the sciatic nerve and optic nerve produced a range of neurotrophins including nerve growth factor (NGF), brain derived neurotrophic factor (BDNF) and neurotrophin-3 (NT-3).

RESULTSThe effects of sciatic nerve and optic nerve CM on neurite outgrowth were tested against a range of neurotrophins, and they had different neuritogenic activities. Only NGF and sciatic nerve CM had obvious neuritogenic activities, although the concentration of NGF in the sciatic nerve CM was very low.

CONCLUSIONSOur experiment showed that sciatic nerve CM had a higher neurotrophic activity on PC12 cells than optic nerve CM. These results suggested that peripheral nervous system (PNS) and central nervous system (CNS) had different expression levels of neurotrophin, which may in part explain the lack of ability to regenerate the CNS.

Animals ; Brain-Derived Neurotrophic Factor ; genetics ; pharmacology ; Cell Differentiation ; drug effects ; Culture Media, Conditioned ; metabolism ; pharmacology ; Nerve Growth Factor ; genetics ; pharmacology ; Neurotrophin 3 ; genetics ; pharmacology ; Optic Nerve ; metabolism ; PC12 Cells ; cytology ; drug effects ; Rats ; Reverse Transcriptase Polymerase Chain Reaction ; Sciatic Nerve ; metabolism

Objective To construct luciferase reporter plasmids of truncated fragments of different lengths of human guanylate binding protein 5(GBP5)gene promoter and analyze the transcriptional activity of each fragment to determine the core regulatory region.Methods GBP5promoter sequence was amplified by PCR,truncated into five fragments of different lengths and connected to pGL3-basic plasmid.The constructed recombinant plasmids pGL3-GBP5-11/21/31/41/51were transfected into 293FT cells and detected for luciferase activity.The binding sites of transcription factors in GBP5promoter region were predicted by JASPAR software,and Yin-Yang transcription factor 1(YY1)targeting the core regulatory region was selected and verified for the transcriptional regulatory activity.The CDS sequence of YY1 was amplified by PCR to construct the overexpression plasmid pIRES2-EGFP-YY1,which was then co-transfected to 293FT cells with plasmids pGL3-GBP5-21(-1 623 ～ +47 bp)and internal reference plasmid pRL-CMV,and detected for luciferase activity to analyze the regulation of transcription factor YY1 on GBP5 promoter activity.Results Colony PCR and double enzyme digestion identification proved that the plasmid of human GBP5 promoter reporter gene was correctly constructed;JASPAR software predicted that there were multiple transcription factor binding sites such as STAT1,YY1 and Foxp3 in GBP5promoter region.Double luciferase activity assay showed that pGL3-GBP5-21(-1 623 ～ +47 bp)showed the highest promoter activity,while the promoter activity of pGL3-GBP5-41(-520 ～ +47 bp)decreased significantly,suggesting that the core region of GBP5 promoter was located at upstream-1 623 ～-520 bp of 5 'UTR;Overexpression of YY1 significantly activated the GBP5 promoter activity and regulated the expression of GBP5.Conclusion The core regulatory region of human GBP5 promoter was located in upstream-1 623 ～-520 bp of the 5 'UTR,with a binding site of transcription factor YY1 existing in this region.Meanwhile,overexpression of YY1 significantly effected the activity of GBP5 promoter.

The purpose of this study was to investigate the effects of chloroquine diphosphate on the proliferation and apoptosis of human leukemic K562 cells, and to elucidate its possible mechanism of activity. The inhibitory effect of chloroquine diphosphate with different concentrations on K562 cell proliferation was detected by MTT method. Apoptosis was measured by flow cytometry (FCM); morphological analysis of apoptosis was performed after staining with propidium iodide (PI) under fluorescence microscope; cell apoptosis was assessed by the DNA ladder shown agarose gel electrophoresis. After treatment with chloroquine diphosphate, K562 cells were stained by Rhodamine 123 to detect changes in mitochondrial transmembrane potential (DeltaPsim) by FCM. The results showed that the cell viability decreased in dose-dependent manner, following chloroquine diphosphate treatment at different concentrations (1.5625, 3.125, 6.25, 12.5, 25, 50 and 100 micromol/L) for 24, 48 and 72 hours. By FCM analysis, the significant increases of sub-G(1) were observed. DNA ladder was detected and apoptotic nuclei were observed. DeltaPsim decreased in K562 cells after chloroquine diphosphate treatment. It is concluded that the chloroquine diphosphate can inhibit the proliferation of K562 cells and induce cell apoptosis, which may relate to down-regulation of mitochondrial transmembrane potential (DeltaPsim).
Antineoplastic Agents ; pharmacology ; Apoptosis ; drug effects ; Cell Proliferation ; drug effects ; Chloroquine ; analogs & derivatives ; pharmacology ; Dose-Response Relationship, Drug ; Down-Regulation ; drug effects ; Humans ; K562 Cells ; Membrane Potentials ; drug effects ; Mitochondria ; drug effects

OBJECTIVETo investigate the possibility of bridging small peripheral nerve gap using a de-acetyl chitosan conduit.

METHODSThe sciatic nerves of right sides were cut at SD rats. They were divided into 5 Groups randomly; Group A: epineurium suture in situ (n = 24); Group B: biological conduit with a small gap for bridging the peripheral nerve (n = 24, with 5 mm gap); Group C: epineurium suture with distal stump rotated 180 degrees (n = 24); Group D: bridging the nerve by biological conduits with a small gap, but the distal stump rotated 180 degrees (n = 24, with 5 mm gap); Group E: biological conduit with a small gap for bridging the peripheral nerve with NGF (n = 24). Electrophysiological examination, histological examination and myelinated axon counting were applied after 2, 4, 6, 8 weeks after operation respectively.

RESULTSRegenerated nerve fibers were seen in the distal nerve segments of all 5 groups; The nerve conduction velocity of small gap group (group B, D) was faster than that of corresponding simple epineurium suture group (group A, C) at all 2, 4, 6, 8 week time point (P < 0.05). The myelinated axon counting of small gap group (group B, D) was faster than that of corresponding simple epineurium suture group (group A, C) at all 4, 6, 8 week time point (P < 0.01), and there was no statistically significant difference at 2 week time point.

CONCLUSIONThe repair effects of chitin conduit bridging peripheral nerve with small gap (5 mm) are better than that of epineurium suture directly, and possess the potential to substitute the epineurium suture.

Animals ; Biocompatible Materials ; Chitosan ; Male ; Nerve Regeneration ; Neurosurgical Procedures ; instrumentation ; methods ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Sciatic Nerve ; injuries ; physiology ; surgery

OBJECTIVETo study the effect of suspension exercise training on motor and balance functions in children with spastic cerebral palsy.

METHODSA total of 97 children with spastic cerebral palsy were randomly divided into an observation group with 49 children and a control group with 48 children. Both groups were given routine rehabilitation training, and the children in the observation group were given suspension exercise training in addition. The scores of the D and E domains of the 88-item version of the Gross Motor Function Measure (GMFM-88) and Berg Balance Scale (BBS) were recorded before treatment and at 1, 3, and 6 months after treatment. Surface electromyography was performed to observe the changes in the root mean square (RMS) of surface electromyogram signals of the adductor muscle and the gastrocnemius muscle.

RESULTSOver the time of treatment, both groups had varying degrees of improvement in the scores of the D and E domains of GMFM-88 and BBS. Compared with the control group, the observation group had significantly greater improvements in D and E functional areas and balance function (P<0.05). Both groups had reductions in the RMS of the surface electromyogram signals of the adductor muscle and the gastrocnemius muscle over the time of treatment, and the observation group had significantly greater reductions than the control group (P<0.05).

CONCLUSIONSSuspension exercise training can effectively improve the motor and balance functions of children with spastic cerebral palsy.

Cerebral Palsy ; physiopathology ; therapy ; Child ; Child, Preschool ; Exercise ; Female ; Humans ; Male ; Motor Activity ; Muscles ; physiopathology

Objective To understand the situation of congenital defects' in five counties/cities in Gansu province so as to provide scientific evidence for the development of effective interventions.Methods General imformaton was collected on all the neonates who were born in Dunhuang city,Jingchuan county,Hui county,Weiyuan county and Yongjing county in Gansu province between Oct.1st,2009 to Sep.304th,2010,with all of their gestational age above 28 weeks.Neonates would include live birth,dead fetus and still birth.Results The overall incidence of congenital defects was 7.49‰ in the five counties/cities in Gansu province in 2009.Ranking order in the top three showed as congenital heart disease,pigmented nevus and limb deformity.Disease with the highest mortality was congenital heart disease (0.79‰).The incidence of congenital defects was 8.35‰ in 2010 with the ranking order of the top three as congenital heart disease,neural tube defects/pigmented nevus and hydrocephalus.Diseases having the highest mortality was congenital heart disease (1.10‰o).Different incidence rates on congenital defects were seen in realted areas,with the highest incidence as 14.65‰ in Dunhuang city.Hui county had the lowest incidence—3.28‰.Conclusion Different incidence of congenital defects were seen in respective areas in Gansu province,with the change of ranking orders.Different strategies should be developed differently depending on the current states of congenital defects in respective areas,according to the three-grade prevention model,to reduce the occurrence of congenital defects.