Objective To screen for differential expression of genes in bone marrow mesenchymal stem ceils (MSCs) stimulated by electromagnetic fields (EMFs) and to study the mechanism of differentiation of the bone mar- row MSCs induced by EMFs.Methods Mouse bone marrow MSCs were isolated and cultured in vitro.The third- passage cells were stimulated by EMFs,then the total RNA was extracted,purified to cDNA and reversely transcribed to yield a cRNA probe.The cRNA probes from stimulated and control cells were labelled with Cy5 and Cy3,respec- tively.The two samples were hybridized with a mouse oligo microarray,and the hybridization signals were scanned. The reverse transcriptase polymerase chain reaction (RT-PCR) was used to identify the interesting genes:Duspl and Rabl.Results A total of 153 differentially expressed genes were found comparing the stimulated group and the control group.There were 74 up-regulated genes and 79 down-regulated genes in the stimulated group.Semi-quantita- tive RT-PCR confirmed that the Duspl and Rabl mRNA expression levels of the stimulated group were up-regulated. Conclusion The gene expression profiles of the bone marrow MSCs were altered after EMF exposure.The genes are involved in cell proliferation and differentiation,transcription control,metabolism and response to stress.These re- sults provide deeper insight into the mechanism of differentiation of the bone marrow MSCs.

Objective To study the effects of 50 Hz magnetic fields on the in vitro proliferation of human osteosareoma cell line MG-63 with different cell densities.Methods Four different magnetic intensities(1 mT, 2 mT,3 roT,4 mT)were used to stimulate the cells,and the experiment was repeated with different cell densities. The method of MTT was employed to evaluate the level of proliferation.Results Fifty Hz magnetic fields signifi- cantly affected the level of proliferation of human osteosareoma cell line MG-63,and the 2 mT intensity exerted the greatest influence on it.The effects of the magnetic field differed with different cell densities.Conclusion The effect of 50 Hz magnetic fields on the in vitro proliferation of human osteosarcoma cell line MG-63 was not only relat- ed to the magnetic intensity,but also the cell density,

OBJECTIVETo establish discriminant functions of diarrhea-predominant irritable bowel syndrome (IBS-D) by studying it from quantitative diagnosis angle, hoping to reduce interference of subjective factors in diagnosing and differentially diagnosing Chinese medical syndromes of IBS-D.

METHODSA Chinese medical clinical epidemiological survey was carried out in 439 IBS-D patients using Clinical Information Collection Table of IBS. Initial syndromes were obtained by cluster analysis. They were analyzed using step-by-step discrimination by taking information of four Chinese medical diagnostic methods and serum brain-gut peptides (BGP) as variables.

RESULTSClustering results were Gan stagnation Pi deficiency syndrome (GSPDS), Pi-Wei weakness syndrome (PWWS), Gan stagnation qi stasis syndrome (GSQSS), Pi-Shen yang deficiency syndrome (PSYDS), Pi-Wei damp-heat syndrome (PWDHS), cold-damp disturbing Pi syndrome (CDDPS). Of them, GSPDS was mostly often seen with effective percentage of 34. 2%, while CDDPS was the least often seen with effective percentage of 5.5%. A total of 5 discriminant functions for GSPDS, PWWS, GSQSS, PSYDS, and PWDHS were obtained by step-by-step dis- crimination method. The retrospective misjudgment rate was 4.1% (16/390), while the cross-validation misjudgment rate was 15.4% (60/390).

CONCLUSIONThe establishment of discriminant functions is of value in objectively diagnosing and differentially diagnosing Chinese medical syndromes of IBS-D.

Alarmins ; Brain ; Cluster Analysis ; Diarrhea ; classification ; diagnosis ; Hot Temperature ; Humans ; Irritable Bowel Syndrome ; classification ; diagnosis ; Medicine, Chinese Traditional ; Qi ; Retrospective Studies ; Surveys and Questionnaires ; Yang Deficiency

AIM: To observe the change of subunit of NADPH oxidation enzyme complex nox - 1 protein in cardiocyte hypoxia - reoxygenation injury and the role of cardiotrophin -1.METHODS: Cardiomyocytes from the hearts of 1 -3 d old neonatal rats were prepared by a modified method. Five groups were included in the study: control; hypoxia/ reoxygenation; hypoxia/reoxygenation + CT - 1; CT - 1 + hypoxia/reoxygenation + LY294002 (PIK3/Akt inhibitor) ; CT -1 + hypoxia/reoxygenation + PD98059 (ERK inhibitor) ; CT - 1 + hypoxia/reoxygenation + DMSO. The concentration of CT -1 was 10 μg/L. The survival rate of myocytes was evaluated by MTS method. Apoptosis, mitochondrial permeability transition pore ( △ψm) and reactive oxygen species ( ROS) were detected by flow cytometry. Nox - 1 protein was determined by Western blotting. RESULTS: Apoptosis of cardiomyocytes and the level of ROS (19.7% ±1.4% vs 2.1% ± 0.5% , 14.07% ± 1.25% vs 3.54% ± 0.86% , P < 0.05 ) increased markedly after hypoxia/reoxygenation, but cardio-myocyte survival rate and the level of△ψm (40.55% ±4.25% vs 86.28% ±7.15% , P <0.01) decreased significantly. The expression of nox - 1 protein was upregulated markedly. With CT - 1 intervention, cardiomyocyte survival rate increased markedly, apoptosis, both ROS and expression of nox - 1 protein reduced significantly. The level of△ψm increased obviously. The effect of CT - 1 was inhibited by LY294002.No significant effect was observed on cells survival in DMSO group, which confirmed that LY294002 was specifically involved in blocking the protective effect of CT - 1.CONCLUSION : The expression of subunit of NADPH oxidation enzyme complex nox - 1 protein is upregulated markedly in cardiocyte hypoxia - reoxygenation injury.CT - 1 protects cardiac cells against hypoxia - reoxygenation injury by downregulating the expression of nox -1 protein to decrease the level of ROS.

Objective To study the effect of Cardiotrophin-1 (CT-1) on cardiocyte hypoxia-reoxygenation injury,and to investigate the signaling pathways involved in the protective effect. Method This study was carried out in Key Lab of Molecular Medicine in Jiangxi Province. Cardiomyocytes from the hearts of 2-day-old Sprague-Dawley neonatal rats were prepared by a modified method. Five groups were included in the study. Group (ⅰ): control, Group (ⅱ): hypoxia/reoxygeuation, Group (ⅲ): hypoxia / reoxygenation + CT-1, Group (iv) : CT- 1 + hypoxia/ reoxygenation + LY294002 (PIK3/Akt inhibitor), Group (ⅴ): CT-1 + hypoxia / reoxygenation +DMSO. The concentration of CT-1 was 10 ng/mL. Myocytes survival rote was evaluated by MTS method, apopto-sis, mitochondrial permeability transition pore (△ψm) and reactive oxygen species(ROS) were detected by flow cy-tometer, phosphorylased GSK-3β and PI3K protein by western blotting. Analysis of variance and q test as statistical methods was used to analyze the data. Results Cardiomyocyte apoptosis and ROS increased markedly after hy-poxia/reoxygenation,but cardiomyocyte survival rate and the level of △ψm [(40.55±4.25) vs. (86.28±7.15), P < 0.01]decreased significantly. With CT-1 intervention, cardiomyocyte survival rate increased markedly (87%),apoptosis and ROS reduced significantly. The level of △ψm increased, the level of phosphorylased GSK-3β and phosphorylased PI3K protein obviously increased. The effect of CT-1 was inhibited by LY294002, but no significant effect was observed on ceils survival in DMSO group, which confirmed that LY294002 specifically in-volved blocking the protective effect of CT-1. Conclusions CT-1 can protect cardiac cells against hypoxia- reoxy-genation injury, these effects are dependent upon its ability to activate the PI3K/GSK-3β pathway.

China ; epidemiology ; Humans ; Rickettsia Infections ; epidemiology ; Seroepidemiologic Studies

OBJECTIVETo summarize the experience and short-term clinical outcomes of hand-assisted laparoscopic surgery (HALS) in sphincter-preserving surgery for low and ultralow rectal cancer.

METHODSData of 49 patients with rectal cancer who underwent HALS for low or ultralow anterior resection between January 2010 and January 2011 were analyzed retrospectively.

RESULTSThe proximal resection margin was (14.3±6.9) cm and the distal margin was(4.3±1.9) cm. The mean operative time was(128.3±70.9) min. On postoperative macroscopic evaluation, the mesorectum was intact in 42 cases, nearly intact in 7 cases. The circumferential resection margin was more than 2 mm in 42 cases, and less than 2 mm in 7 cases. Forty-six patients underwent R0 resection, and 3 cases underwent R1 resection. The median retrieved lymph node (LN) was 16.20±9.23, and the median positive LN was 1.12±2.19. Postoperative pathological examination showed TNM stage was I( in 12 patients, II(A in 18, II(B in 1, III(A in 2, III(B in 8, III(C in 5, IIII( in 3. The median postoperative hospital stay was (6.25±3.87) d. There were no anastomotic leakage, ileus, intra-abdominal or anastomotic bleeding. There were two wound infections.

CONCLUSIONLow and ultralow anterior resection for rectal cancer using HALS approach is safe and feasible with favorable short-term outcome.

Adult ; Aged ; Aged, 80 and over ; Anal Canal ; surgery ; Female ; Hand-Assisted Laparoscopy ; methods ; Humans ; Male ; Middle Aged ; Rectal Neoplasms ; surgery ; Retrospective Studies ; Treatment Outcome

Chemical constituents from the acetone extract of twigs of Manglietia hookeri were isolated and purified by various column chromatographic methods over silica gel and sephadex LH-20, and preparative TLC. The structures of these compounds were identified on the basis of physicochemical properties and spectral analysis, including NMR and MS spectra. Six eudesmane sesquiterpenes were obtained and their structures were identified as trans-eudesmane-4, 11-diol(1), β-eudesmol(2), (-) -10-epi-5β-hydroxy-β-eudesmol (3), epi-carrisone (4), 6-hydroxy-eudesm-4(14) -ene(5) and gynurenol(6). All the compounds were isolated from this plant for the first time. Furthermore, the 13C-NMR data of compound 3 were reported for the first time.
Drugs, Chinese Herbal ; chemistry ; isolation & purification ; Magnetic Resonance Spectroscopy ; Magnolia ; chemistry ; Molecular Structure ; Plant Stems ; chemistry ; Sesquiterpenes, Eudesmane ; chemistry ; isolation & purification ; Spectrometry, Mass, Electrospray Ionization

OBJECTIVETo find out potential molecular targets for gallbladder carcinoma diagnosis and treatment by analyzing and comparing the proteins expressed in human gallbladder carcinoma tissue and benign gallbladder tissue.

METHODSProteomic analysis of 6 human gallbladder carcinoma tissues and 6 benign gallbladder tissues was carried out. Total proteins of the carcinoma tissue and benign gallbladder tissue were separated by two-dimensional gel electrophoresis (2-DE). The differentially expressed proteins were analyzed and identified by matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF-MS). Immunohistochemistry was used to examine the expression of PEBP1 protein in an independent series of samples.

RESULTSProtein extracts of individual samples in each type of tissues were separated on two-dimensional gels. There were forty six differentially expressed proteins in the gallbladder carcinom tissues. Seventeen proteins were successfully identified by MS, in which nine proteins were overexpressed in tumors while the other eight proteins were underexpressed. The increased level of PEBP1 protein in gallbladder carcinoma was further confirmed by immunohistochemical analysis.

CONCLUSIONSeventeen differentially expressed proteins were successfully characterized by comparative proteomic analysis. Those results may provide scientific foundation for screening the molecular biomarkers which can be used in diagnosis and treatment of gallbladder carcinoma, as well as to improve its prognosis and provide a new clue for carcinogenesis research of gallbladder carcinoma.

Adenocarcinoma ; diagnosis ; metabolism ; pathology ; Adult ; Aged ; Biomarkers, Tumor ; analysis ; Electrophoresis, Gel, Two-Dimensional ; Gallbladder Neoplasms ; diagnosis ; metabolism ; pathology ; Gallstones ; diagnosis ; metabolism ; pathology ; Gene Expression Profiling ; Humans ; Immunohistochemistry ; Middle Aged ; Phosphatidylethanolamine Binding Protein ; metabolism ; Proteomics ; methods ; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization

OBJECTIVETo explore the potential molecular targets for diagnosis and treatment of gallbladder cancer by analyzing and comparing the proteomes expressed in human gallbladder cancer and benign gallbladder tissues.

METHODSThe proteins expressed were analyzed using two-dimensional gel electrophoresis. The differentially expressed proteins in tumors were also analyzed by mass spectrometry (MS). AnnexinA3 expression was examined by streptavidin peroxidase immunohistochemical technique on paraffin-embedded tissue sections from 50 patients of gallbladder cancer and 38 cases of chronic eholecystitis.

RESULTSProtein extracts of individual sample in each type of tissues were separated on two-dimensional gels. There were forty six differentially expressed proteins in the tissue samples of gallbladder cancer. Seventeen proteins were successfully identified by MS, in which nine proteins were overexpressed in tumors and the other eight proteins were underexpressed. The positive expression rates of annexinA3 in gallbladder cancer was significantly higher than that in chronic cholecystitis, and the difference was statistically significant (74.0% vs 21.1%, P < 0.01). In the gallbladder cancer, no correlation was obtained between annexinA3 and age, gender or histologicl type (P > 0.05), but overexpression of annexinA3 correlated significantly with those cases with a lower histological grading (40.0% vs 82.5%, P < 0.05); lymph node or distant metastasis (40.9% vs 100%, P < 0.05); or a shorter survival time after operation (50.0% vs 93.8%, P < 0.05).

CONCLUSIONSSignificant discrepancies in protein expression exist among gallbladder cancer and benign gallbladder tissues. AnnexinA3 plays an important role in the initiation and progression of human gallbladder cancer.

Adenocarcinoma ; metabolism ; pathology ; surgery ; Adult ; Aged ; Annexin A3 ; metabolism ; Carcinoma, Adenosquamous ; metabolism ; pathology ; surgery ; Carcinoma, Squamous Cell ; metabolism ; pathology ; surgery ; Cholecystitis ; metabolism ; Electrophoresis, Gel, Two-Dimensional ; Female ; Gallbladder Neoplasms ; metabolism ; pathology ; surgery ; Gene Expression Profiling ; Humans ; Lymphatic Metastasis ; Male ; Middle Aged ; Neoplasm Metastasis ; Proteome ; metabolism ; Proteomics ; Survival Rate