Objective To investigate the neural proliferation , differentiation and apoptosis of the developing spinal cord of the mouse and to discuss the mechanism of spinal cord ’ s development .Methods 5-Bromodeoxyuridine ( BrdU) assay was used to mark the proliferative neural stem cells , and the immunofluorescent stainings ( DCX, NeuN and Caspase8) were carried out to visualize the newborn neurons , mature cells and apoptotic cells in the spinal cord with 173 mice arrange from E18 to P90.Results BrdU positive neural stem cells appeared evenly in the spinal cord at early days . With age increasing , the neural stem cells differentiated into neuroglial cells and neurons .The newborn neurons in the subventricular zone migrated toward the intermediate zone ( putative gray matter ) and differentiated into mature neurons gradually .With neurons ’ concentrating towards the center , the gray matter formed an “H” shape .In the meantime , with neural differentiation , some apoptotic neurons appeared among the newborn neurons and mature neurons . Double immunostaining showed that most apoptotic neurons were newborn neurons , suggesting the neuroapoptosis more likely occurred in newborn neurons .The statistical data showed that the number of DCX , NeuN and Caspase-8 positive cells reduced with age increasing , suggesting neural differentiation and neuroapoptosis decreased during spinal cord ’ s development .Conclusion Neural proliferation , neural differentiation and neuroapoptosis occur in developing spinal cord . They work together to regulate the formation and development of the spinal cord .

Objective To evaluate the left ventricular transmural mechanics changes of breast cancer patients between before and after anthracycline chemotherapy by two-dimensional speckle tracking imaging (2D-STI),and to predict early cardiotoxicity caused by anthracycline.Methods Forty-six breast cancer patients with postoperative anthracycline-based chemotherapy were recruited.Echocardiography were performed on all subjects before and at 1 ,3 and 6 anthracycline-based chemotherapeutic cycle.Global longitudinal strain(GLS),endocardial longitudinal strain(LS-endo),epicardial longitudinal strain(LS-epi), global radial strain (GRS),endocardial radial strain (RS-endo ),epicardial radial strain (RS-epi ),global circumferential strain (GCS),endocardial circumferential strain (CS-endo)and epicardial circumferential strain(CS-epi) were assessed by 2D-STI and transmural myocardial strain gradient-longitudinal strain (TMSG-LS),transmural myocardial strain gradient-radial strain(TMSG-RS),transmural myocardial strain gradient-circumferential strain(TMSG-CS)were calculated.Conventional echocardiographic parameters and strain-related parameters before and after chemotherapy were compared. The receiver operating characteristics(ROC)curve was performed to determine sensitivity and specifity of strain parameters for prediction value of cardiotoxicity induced by anthracycline chemotherapy.Results ①After the sixth cycle of anthracycline chemotherapy,9 patients (16.4%)had developed anthracycline-induced cardiotoxicity,and 37 patients (80.4%)did not meet the criteria for cardiotoxicity.② There were no significant differences in conventional echocardiography parameters between before and after chemotherapy (P > 0.05 ).Left ventricular ejection fraction (LVEF),fractional shortening (FS)and E/A significantly decreased,but E/e significantly increased after six cycles of chemotherapy (P < 0.05 ).③ When comparison with baseline cases,GLS,LS-endo,LS-epi,TMSG- LS,TMSG-RS decreased after 3,6 cycles of chemotherapy,GRS, RS-endo and RS-epi decreased after six cycles of chemotherapy,the difference was statistically significant (P <0.05 ).④ ROC curve results showed the value of TMSG-LS,LS-endo,GLS and LS-epi after three cycles of chemotherapy to detect of anthracycline-induced cardiotoxicity,the areas under the curve were TMSG-LS> LS-endo> GLS> LS-epi.Conclusions After three cycles of chemotherapy,the decreases of TMSG-LS,LS-endo,GLS and LS-epi preceded the change of LVEF and other strain parameters,TMSG-LS and LS-endo can accurately and early detect anthracycline chemotherapy-induced cardiotoxicity.

A water-soluble polysaccharide fraction from root of Potentilla anserine was obtained. Gas chromatogram, FT-IR, physical and chemical characteristics of the Potentilla anserine polysaccharide fraction (PAPF) were analyzed. The protective effects of PAPF against the H2O2 induced process of apoptosis of murine splenic lymphocytes were investigated in vitro. Morphological assessment of apoptosis was performed with light microscope and laser scanning confocal microscope. DNA fragmentation was visualized by agarose gel electrophoresis. The amount of apoptotic cells was measured by flow cytometry. The results showed that PAPF is composed of rhamnose, arabinose glucose and galactose. H2O2 (200 micromol x L(-1)) induced apoptosis of murine splenic lymphocytes with the cell volume reduced, cytoplasm and nuclear shrunk and DNA stained non-uniformly. Condensed chromatin and formation of apoptotic body were observed in the apoptotic cells. Apoptotic bodies in the cells treated with PAPF and H2O2 were less than those in H2O2 treatment alone. DNA fragmentation assay showed that PAPF (50, 100, 200, and 400 microg x mL(-1)) obviously reduced H2O2-induced ladder bands. Flow cytometry analysis showed that H2O2 increased the populations of apoptotic sub-G1 cells from 5.60% (control) to 45.40%, and PAPF decreased H2O2-induced apoptosis to 37.80%, 22.70%, 17.70%, and 8.50%, respectively. In conclusion, PAPF reduced H2O2-induced oxidative damage in a dose dependent manner.
Animals ; Apoptosis ; drug effects ; Cells, Cultured ; DNA Fragmentation ; Flow Cytometry ; Hydrogen Peroxide ; pharmacology ; Lymphocyte Count ; Lymphocytes ; cytology ; drug effects ; Mice ; Polysaccharides ; pharmacology ; Potentilla ; Spleen ; cytology

AIM:To investigate the effects of exosomes secreted by pancreatic cancer cells on the viability and function of βcells and the possible mechanism .METHODS:ExoQuick-TC kit was used to extract exosomes in the super-natants of mouse pancreatic cancer Pan 02 and MPC-83 cells, and the extracted exosomes were identified by transmission electron microscopy.Fluorescence-labeled exosomes were incubated with mouse insulinoma MIN 6 cells for 48 h to detect whether exosomes secreted by pancreatic cancer cells were uptaken by MIN 6 cells.MTT and glucose-stimulated insulin se-cretion ( GSIS) assays were conducted to examine cell viability and insulin secretion of MIN 6 cells after incubating with ex-osomes.The expression of miR-204 and Bcl-2 mRNA in MIN6 cells was detected by qPCR .The protein expression of Bcl-2, Bax, caspase-3 and cytochrome C (Cyt-C) in MIN6 cells was determined by Western blot .RESULTS:The results of transmission electron microscopy showed that both Pan 02 cells and MPC-83 cells secreted exosomes , and Pan02 cells secre-ted more.The co-incubation results of fluorescence-labeled exosomes and MIN6 cells confirmed that MIN6 cells were able to ingest large amounts of exosomes secreted by pancreatic cancer cells .The results of MTT and GSIS assays showed that the viability and the level of high glucose-stimulated insulin secretion of MIN 6 cells in exosome treatment group significantly decreased compared with nontreatment group (P<0.01).The results of qPCR showed that the exosomes secreted by pan-creatic cancer cells were rich in miR-204, and the mRNA expression of Bcl-2 in MIN6 cells was significantly down-regula-ted by exosome incubation ( P<0.01) .The results of Western blot showed that the protein expression of Bcl-2 in the MIN6 cells treated with exosomes was significantly down-regulated (P<0.05), and the protein levels of Bax, cleaved caspase-3 and Cyt-C in exosomes treatment group were significantly up-regulated ( P<0.01 ) .CONCLUSION: Pancreatic cancer cells secrete exosomes .The exosomes secreted by pancreatic cancer cells are ingested by βcells, and reduce the viability and insulin secretion of βcells.The mechanism may be related to the increase in exosomal miR-204 in the βcells.In-creasing miR-204 may inhibit the expression of Bcl-2 and promote the activation of mitochondrial apoptosis in βcells.

Objective To investigate the predictors of long-term remission of type 2 diabetes induced by short-term intensive insulin treatment.Methods Fifty-four cases of diabetes mellitus with the duration of illness less than 5 years received an intensive insulin treatment for 2 weeks.The standard meal test and intravenous glucose tolerance test were performed at the baseline and 24 h after treatment completion respectively.Long-term remission meant that the diabetic patients should maintain the target glyeaemic control without any hypoglyeaemie agent within one year.Results The remission rate was 57.4% (31/54) overall,and even reached to 80.6% (29/36) in patients with the duration of illness less than 6 months,whereas,the remission rate was only 11.1% (2/18) in those with the duration of illness more than 12 months.In another view,the remission rate was significantly higher in the patients with fasting plasma glucose (FPG) level of less than 7 mmol/L (78.8%,26/ 33) 24 h after intensive treatment than those with FPG level of more than 7 mmol/L (23.8%,5/21,P

To investigate 1alpha,25-(OH)2D3 regulation of matrix metalloproteinase-9 (MMP-9) protein expression during osteoclast formation and differentiation, receptor activator of nuclear factor kappaB ligand (RANKL) and macrophage colony-stimulating factor (M-CSF) were administered to induce the differentiation of RAW264.7 cells into osteoclasts. The cells were incubated with different concentrations of 1alpha,25-(OH)2D3 during culturing, and cell proliferation was measured using the methylthiazol tetrazolium method. Osteoclast formation was confirmed using tartrate-resistant acid phosphatase (TRAP) staining and assessing bone lacunar resorption. MMP-9 protein expression levels were measured with Western blotting. We showed that 1alpha,25-(OH)2D3 inhibited RAW264.7 cell proliferation induced by RANKL and M-CSF, increased the numbers of TRAP-positive osteoclasts and their nuclei, enhanced osteoclast bone resorption, and promoted MMP-9 protein expression in a concentration-dependent manner. These findings indicate that 1alpha,25-(OH)2D3 administered at a physiological relevant concentration promoted osteoclast formation and could regulate osteoclast bone metabolism by increasing MMP-9 protein expression during osteoclast differentiation.
Acid Phosphatase/metabolism ; Animals ; Blotting, Western ; Calcitriol/*pharmacology ; Calcium Channel Agonists/pharmacology ; *Cell Differentiation ; Cell Line ; Cell Proliferation ; Gene Expression Regulation, Enzymologic/*drug effects ; Isoenzymes/metabolism ; Matrix Metalloproteinase 9/*genetics/metabolism ; Mice ; Osteoclasts/*cytology/*enzymology ; Tetrazolium Salts ; Thiazoles

Objective of this study was to perform bioinformatics analysis of the characteristics of gene expression profiling regulated by amifostine and predict its novel potential biological function to provide a direction for further exploring pharmacological actions of amifostine and study methods. Amifostine was used as a key word to search internet-based free gene expression database including GEO, affymetrix gene chip database, GenBank, SAGE, GeneCard, InterPro, ProtoNet, UniProt and BLOCKS and the sifted amifostine-regulated gene expression profiling data was subjected to validity testing, gene expression difference analysis and functional clustering and gene annotation. The results showed that only one data of gene expression profiling regulated by amifostine was sifted from GEO database (accession: GSE3212). Through validity testing and gene expression difference analysis, significant difference (p < 0.01) was only found in 2.14% of the whole genome (460/192000). Gene annotation analysis showed that 139 out of 460 genes were known genes, in which 77 genes were up-regulated and 62 genes were down-regulated. 13 out of 139 genes were newly expressed following amifostine treatment of K562 cells, however expression of 5 genes was completely inhibited. Functional clustering displayed that 139 genes were divided into 11 categories and their biological function was involved in hematopoietic and immunologic regulation, apoptosis and cell cycle. It is concluded that bioinformatics method can be applied to analysis of gene expression profiling regulated by amifostine. Amifostine has a regulatory effect on human gene expression profiling and this action is mainly presented in biological processes including hematopoiesis, immunologic regulation, apoptosis and cell cycle and so on. The effect of amifostine on human gene expression need to be further testified in experimental condition.
Amifostine ; pharmacology ; Computational Biology ; Gene Expression ; drug effects ; Gene Expression Profiling ; methods ; Humans ; Microarray Analysis ; Molecular Sequence Annotation

OBJECTIVETo observe the effect of inhalation of aerosolized perfluorocarbon combined with tetramethylpyrazine on the hemodynamics and histopathology in a porcine model of acute lung injury.

METHODSNormal adult pigs were subjected to saline lavage of the bilateral lungs to induce acute lung injury and randomized subsequently into 3 groups for treatment with inhalation of perfluorocarbon, combined inhalation of perfluorocarbon and tetramethylpyrazine, or inhalation of tetramethylpyrazine. The changes of mean arterial pressure (MAP), PetCO(2), mPAP, CVP and PAWP were recorded at different time points following the lung injury, and the lung tissues were sampled for histological observations.

RESULTSThe MAP, mPAP, CVP and PAWP all increased significantly in the 3 groups after acute lung injury. Interventions with combined tetramethylpyrazine and perfluorocarbon inhalation significantly improved these indices as compared with inhalation of tetramethylpyrazine or perfluorocarbon alone (P<0.05). The pulmonary pathology was the mildest in the combined inhalation group, and the most severe in tetramethylpyrazine group.

CONCLUSIONCombined inhalation of perfluorocarbon and tetramethylpyrazine can effectively improve the oxygenation, reduce pulmonary arterial pressure?and ameliorate lung pathology in pigs with acute lung injury.

Acute Lung Injury ; drug therapy ; etiology ; pathology ; Administration, Inhalation ; Animals ; Drug Therapy, Combination ; Fluorocarbons ; administration & dosage ; Hemodynamics ; drug effects ; Lung ; pathology ; Phytotherapy ; Pyrazines ; administration & dosage ; Swine

OBJECTIVETo develop and evaluate the efficiency of air purification and sterilization instrument based on nano-sized TiO(2) photocatalytic technique.

METHODSThe nano-sized TiO(2) photocatalytic air purification and sterilization instrument was designed and a sample had been prepared. The sterilization efficiencies for E.coli and Klebsiella by the nano-sized TiO(2) photocatalytic instrument and ultraviolet (UV) were measured in closed labs. The on-site efficiency of the instrument was evaluated, too.

RESULTSThe nano-sized TiO(2) photocatalytic air purification and sterilization instrument was composed of five units: rough filter, nano-sized TiO(2) photocatalytic unit, activated carbon fiber filter, negative ion generator, and programmed control unit. The E.coli killing rates by the nano-sized TiO(2) photocatalytic instrument were 76.0%, 81.8%, 77.5%, and 80.7% at 30, 60, 90, and 120 minutes, respectively. There was no significant difference between the E.coli killing rates of the instrument and UV (P > 0.05), except the 120 minutes timepoint. The Klebsiella killing rates by the instrument were 78.4%, 79.5%, 67.3%, and 58.5% at 30, 60, 90, and 120 minutes, respectively. The Klebsiella killing efficiencies of the instrument at 30 and 60 minutes were better than that of UV (P < 0.01). There was no significant difference between the Klebsiella killing efficiencies of the instrument and UV (P > 0.05).

CONCLUSIONThe air sterilization efficiency of the nano-sized TiO(2) photocatalytic instrument should be equivalent or better as compared with the UV. This instrument might be used for the air purification and sterilization of the public locations.

Air Pollution ; prevention & control ; Decontamination ; methods ; Disinfection ; methods ; Nanostructures ; Photochemistry ; Titanium

OBJECTIVETo study the significance of supratubal recess and its aeration pathway to epitympanum in the pathogenesis of cholesteatoma otitis media.

METHODSFifty-two ears of cholesteatoma were selected as study group. Sixteen ears of traumatic facial palsy with pneumatic mastoid, which had no history of chronic otitis media were selected as control group. The status of supratubal recesses of all and their aeration pathways to epitympanum were observed in operations.

RESULTSSixteen ears from control group clearly presented supratubal recesses. Membrane closure was founded in four of them. The aeration pathways of fifty-two ears (100%) from study group were all completely closed. Comparing with control group, the difference was obviously significant (chi2 = 41.7144, P = 0.000). Among these cases, bony closure was observed in thirty-four ears (65.4%), while membrane closure in eighteen ears (34.6%). Their epitympanum space was very narrow and mastoid was sclerotic or poorly developed.

CONCLUSIONSBlockage of the aeration pathway between supratubal recess and epitympanum was possible one of the origins of negative-pressure status of epitympanum and mastoid, which might lead to the formation of aural cholesteatoma. Anatomy variation of the aeration pathway from supratubal recess to epitympanum might be a pathogenesis factor of cholesteatoma otitis media. It suggested that opening the aeration pathway in tympanoplasty with intact canal wall up technique might be helpful to prevent recurrence of aural cholesteatoma.

Adolescent ; Adult ; Aged ; Case-Control Studies ; Cholesteatoma, Middle Ear ; pathology ; Eustachian Tube ; pathology ; Facial Paralysis ; pathology ; Female ; Humans ; Male ; Middle Aged ; Young Adult