BACKGROUND: Umbilical cord blood-mesenchymal stem cells (UCB-MSCs) following differentiation into cardiomyocytes were transplanted into ischemic myocardium. The transplanted cells can build connection with host cells and repair the infarct myocardium. OBJECTIVE: To detect the cell-cell junction after transplantation of the cardiac-like cell derived from the canine umbilical cord blood stem cells. DESIGN, TIME AND SETTING: A randomized controlled animal study was performed from July 2006 to October 2007 at the Animal Experimental Center of Xinhua Hospital Affiliated to School of Medicine, Shanghai Jiao Tong University. MATERIALS: A total of 2 full-term pregnant canines were used for isolation of UCB-MSCs. A total of 36 adult mongrel canines were divided into cell transplantation group and model control group (n=18) according to the rule of random digits table. METHODS: The MSCs at passage 4 were transfected by Laz-Z. After 3-day culture, MSCs were induced by 10 μmol/L 5-azacytidine (5-aza). The canine models of myocardium infarction were established following 3 weeks of culture. 2 mL (1 ×107)MSCs were transplanted into dogs with acute myocardium infarction by coronary artery infusion and local injection in cell transplantation group. An equal volume of saline was used in the model control group. The specimens were harvested and detected at 2, 4 and 8 weeks, respectively. Cell junction was determined using immunohistochemistry. MAIN OUTCOME MEASURES: The following parameters were measured: gene trensfection, myogenic induction and differentiation results of UCB-MSCs; junction of transplanted cells and host cardiomyocytes. RESULTS: Following 72 hours of transfaction, mass of cells expressed LacZ gene, synthetized galactosidase, and stained blue using X-gal staining. Following 3 weeks of 5-aza induction, the antigen a-Actin, Desmin and Connexin43 were all been positively expressed, but before induction they were all negative. From the myocardial section of 8 weeks after transplantation, the junction was formed between the transplanted cells and the host myocardium as formed between the transplanted cells. In the junction, green-fluorescence positive expression of cadherin and connexin43 could be seen. However, in the model control group, only cadherin and connexin43 expressed positively, but the transplanted UCB-MSCs with red fluorescence could not been observed. CONCLUSION: The UCB-MSCs is able to differentiate into cardiac-like cell in vitro and form cell-cell junction in vivo to communicate with surrounding cells.

OBJECTIVETo investigate the effects of ischemic postconditioning (IPostC) on pneumocyte apoptosis after lung ischemia/reperfusion injury in rats.

METHODSAdult male SD rats were randomly divided into 3 groups based upon the intervention (n = 8): control group (C), lung ischemic reperfusion group (LIR), LIR+ IPostC group (IPostC). At the end of the experiment, blood specimens drawn from the arteria carotis were tested for the content of malondialdehyde (MDA), the activity of superoxide dismutase (SOD) and myeloperoxidase (MPO); the pneumocyte apoptosis index (AI) was achieved by tennrminal deoxynucleotidyl transferase mediated dUTP nick end abeling (TUNEL); the expression of Bcl-2, Bax protein in lung tissue was accessed by quantitative immunohistochemistry (MHC) and Bcl-2, Bax mRNA by RT-PCR.

RESULTSIPostC could significantly attenuate the MDA level, MPO activity and improve SOD activity in blood serum which was comparable to I/R and significantly reduced the number of TUNEL-positive cells compared with I/R group, expressed as Al (% total nuclei) from (39.0 +/- 3.46) to (8.0 +/- 0.88) (P < 0.01). The protein and mRNA expression of Bcl-2 and Bax showed that IPO significantly attenuated the ischemia/reperfusion-upregulated expression of Bax protein but improved the expression of Bcl-2 that improved the Bcl-2/Bax ratio (P < 0.01) .

CONCLUSIONIPostC may attenuate pneumocyte apoptosis in LIRI by up-regulating expression of Bcl-2/Bax ratio and by inhibiting oxidant generation and neutrophils filtration.

Alveolar Epithelial Cells ; cytology ; Animals ; Apoptosis ; Ischemic Postconditioning ; Lung ; metabolism ; pathology ; Lung Injury ; physiopathology ; Male ; Malondialdehyde ; metabolism ; Peroxidase ; metabolism ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; Rats ; Rats, Sprague-Dawley ; Reperfusion Injury ; physiopathology ; Superoxide Dismutase ; metabolism ; bcl-2-Associated X Protein ; metabolism

OBJECTIVETo study a method for extraction and analysis of volatile components from Chrysanthemum morifolium 'gonghuangjv' cv. nov (CM GHJ) and C. morifolium 'gongbaijv' cv. nov (CM GBJ) by solid-phase microextraction (SPME) and gas chromatography-mass spectrometry (GC-MS).

METHODThe volatile components were extracted in different temperature, different balance period and different extraction fiber using head space solid-phase microextraction (HS-SPME), and were identified by CGC-MS. The variety in integral area of each component was observed in different conditions and its relative content was determined by normalization of area.

RESULTThe better condition of SPME for C. morifolium was that the sample was extracted using 100 microm polydimethylsiloxane (PDMS) extraction fiber after it had been balanced for 6 hours at 75 degrees C. 53 components from CM GHJ and CM GBJ were identified, and there were 35 same components in CM GHJ and CM GBJ.

CONCLUSIONHS-SPME-GC-MS is convenient, rapid and reliable for analysis of volatile components in C. morifolium.

Chrysanthemum ; chemistry ; classification ; Flowers ; chemistry ; Gas Chromatography-Mass Spectrometry ; methods ; Oils, Volatile ; analysis ; Plants, Medicinal ; chemistry ; classification

Objective To study the effect of metformin on glucolipid metabolic disorders and liver lipid deposition caused by clozapine in rats.Methods From 1 d to 4 d,Clozapine 5 mg·kg -1 ·d -1 was gavaged,and the dose increased to 25 mg·kg -1 ·d -1 from the 5th day.Metformin 100 mg·kg -1 ·d -1 or 400 mg·kg -1 ·d -1 or simvastatin 1 mg· kg -1 ·d -1 was gavaged from the 15th day.The total period of dosing was 8 weeks.Body mass,fasting blood sugar (FBS) and postprandial 2 hours blood glucose (2hPBG)were measured at baseline,3 d,1 week,2 weeks,4 weeks,6 weeks and 8 weeks.At the end of the 8th week,serum cholesterol (TC),triglyceride (TG),low density lipoprotein (LDL -C), high density lipoprotein (HDL -C),fructosamine (FA)and insulin (IRS)were measured and liver HE staining was done.Results There were no significant differences of the measured indexes between control group and metformin group at the all test points.By the end of the 6th and 8th week,compared with control group,the body mass,FBS,2hPBG,IRS, FA,TC,TG and LDL -C were significantly increased in clozapine group (P <0.05 ),while HDL -C decreased in clozapine group (P <0.05).Compared with clozapine group,body mass,FBS,2hPBG,IRS,FA,TC,TG and LDL -C were significantly decreased by metformin or simvastatin administration (P <0.05),while HDL -C increased(P <0.05).Rat liver cells in clozapine group were not neat around the small blood vessels;there were more white fat cells and hepatocellular lipid calm far away from the blood vessels.However,in other groups,there were moderate white fat cells, and there were not much hepatocellular lipid calm far away from the blood vessels.Conclusion Metformin could effectively prevent and treat weight gain,glucolipid metabolic disorder and liver lipid deposition caused by clozapine.

Objective: Exposure to the herbicide glyphosate during pregnancy and lactation may increase the risk for autism spectrum disorder (ASD) in offspring. Recently, we reported that maternal exposure of formulated glyphosate caused ASD-like behaviors in juvenile offspring. Here, we investigated whether maternal exposure of pure glyphosate could cause ASD-like behaviors in juvenile offspring. Methods: Water or 0.098% glyphosate was administered as drinking water from E5 to P21 (weaning). Behavioral tests such as grooming test and three-chamber social interaction test in male offspring were performed from P28 to P35. Results: Male offspring showed ASD-like behavioral abnormalities (i.e., increasing grooming behavior and social interaction deficit) after maternal exposure of glyphosate. Conclusion The findings suggest that the exposure of glyphosate during pregnancy and lactation may cause ASD-like behavioral abnormalities in male juvenile offspring. It is likely that glyphosate itself, but not the other ingredients, may contribute to ASD-like behavioral abnormalities in juvenile offspring.

Objective: Exposure to the herbicide glyphosate during pregnancy and lactation may increase the risk for autism spectrum disorder (ASD) in offspring. Recently, we reported that maternal exposure of formulated glyphosate caused ASD-like behaviors in juvenile offspring. Here, we investigated whether maternal exposure of pure glyphosate could cause ASD-like behaviors in juvenile offspring. Methods: Water or 0.098% glyphosate was administered as drinking water from E5 to P21 (weaning). Behavioral tests such as grooming test and three-chamber social interaction test in male offspring were performed from P28 to P35. Results: Male offspring showed ASD-like behavioral abnormalities (i.e., increasing grooming behavior and social interaction deficit) after maternal exposure of glyphosate. Conclusion The findings suggest that the exposure of glyphosate during pregnancy and lactation may cause ASD-like behavioral abnormalities in male juvenile offspring. It is likely that glyphosate itself, but not the other ingredients, may contribute to ASD-like behavioral abnormalities in juvenile offspring.

The objective of this study was to estimate a novel apoptosis-promoting molecule PDCD5 expression in the bone marrow cells from adult acute myeloid leukemia (AML) for investigation of its significance in the pathogenesis of AML. Flow cytometry assay was used for detection of PDCD5 expression in the different groups of cells from bone marrow of AML patients and normal controls by using 21 monoclonal antibodies with different fluorescent markers. The PDCD5 expressions in bone marrow cells from some AML patients and normal controls were also detected by Western blot. The results showed that the mean PDCD5 fluorescence intensity in bone marrow nucleated cells (MNC) from the bone marrow of 36 untreated AML patients was significantly lower than that from the bone marrow of 30 normal controls (3059 +/- 1392) vs (7432 +/- 1261) (P < 0.01). The mean PDCD5 fluorescence intensity was lower in the marrow granulocytes, monocytes, blast cells, and lymphocytes from untreated AML patients than that from normal (3939 +/- 2121) vs (8367 +/- 1045); (3156 +/- 1635) vs (5917 +/- 2329); (2824 +/- 1592) vs (3998 +/- 2106); (1474 +/- 816) vs (3355 +/- 2042) respectively, (all P < 0.01). Western blot analysis demonstrated that PDCD5 expression was significantly decreased in the AML cells, as compared with normal cells. It is concluded that PDCD5 expression in MNC in untreated AML patients is lower than that in the normal. PDCD5 expression in the marrow granulocytes, monocytes, blast cells, and lymphocytes of untreated AML patients is significantly lower than that in the normal. It suggests that the abnormally low expression of PDCD5 may be involved in the pathogenesis of AML.
Apoptosis ; physiology ; Apoptosis Regulatory Proteins ; metabolism ; Bone Marrow Cells ; metabolism ; Humans ; Leukemia, Myeloid, Acute ; metabolism ; pathology ; Neoplasm Proteins ; metabolism

Objective To investigate the molecular mechanisms of upregulated expression of cellular Fas-associated death domain-like interleukin-1 beta converting enzyme inhibitory protein(c-FLIP)by calreticulin(CRT)in patients with rheumatoid arthritis (RA). Methods The semi-quantitative analysis and localization of c-FLIP in RA and osteoarthritis (OA)synovium were detected by immunohistochemistry.The fibroblast-like synoviocytes(FLS)were isolated by enzymatic digestion of synovial tissue specimens obtained from RA and OA patients,and cultured as an in vitro experiment model.The expressions of c-FLIP in RA and OA synovial fibroblasts were detected by immunofluorescence and Western blot assay. Whether CRT influenced c-FLIP expression and its molecular mechanism were explored by Western blot assay. Results The high expression of c-FLIP was found in RA synovium, mainly in the lining and sublining areas of FLS and vascular endothelial cells detected by immunohistochemistry.Meanwhile,weak staining of c-FLIP was observed in OA synovium.The expression of c-FLIP was significantly higher in RA synovium than that of OA synovium(t=11.717,P<0.001).Results of immunofluorescence and Western blot assay showed that c-FLIP was mainly located in cytoplasm, and which was higher expressed in FLS of RA than that of OA. The increased c-FLIP expression and phosphorylation of NF-κB were detected after being co-incubated with exogenous CRT (0, 10, 50, 100 μg/L), in dose-dependent manner. The effect of CRT upregulating c-FLIP expression was blocked by NF-κB inhibitor BAY 11-7082.Conclusion CRT can increase c-FLIP expression at least partly through NF-κB pathway in RA,which may provide therapeutic target for the treatment of RA.

BACKGROUND: It is of vital importance to fabricate an interface on the titanium implant surface which can promote early cell adhesion, proliferation, and differentiation, and exert better antibacterial effects with no cytotoxicity. OBJECTIVE: To prepare a TiN/Ag composite coating on the surface of pure titanium implant, and to explore its antibacterial properties and effects on MC3T3-E1 biobehaviors. METHODS: Acid etching blasting and multi-arc ion plating were adopted to prepare TiN/Ag composite coating on the smooth surface of pure titanium. Then, MC3T3-E1 cells that grew well were inoculated onto pure titanium plate, sandblasted and acid-etched titanium plate, and TiN/Ag-coated titanium plate. Twenty-four hours later, cell adhesion and viability were observed under confocal laser scanning microscope, and cell morphology was observed under scanning electron microscope. Cell counting kit-8 was used to detect cell proliferation and cytotoxicity at 24,48,72 hours after inoculation.In addition,Staphylococus aureus solution was dropped onto the smooth titanium plated, acid-etched and sandblasted titanium and TiN/Ag-coated titanium plate, and the growth of bacteria was observed by the laser confocal scanning microscope at 16 hours. RESULTS AND CONCLUSION: Under the confocal laser scanning microscope, spindle cells with bipolar or three poles were observed on the smooth titanium surface, and there was less F-actin and filopodia expression; cells on the TiN/Ag-coated titanium surface and sandblasted and acid-etched titanium surface were scattered with a large amount of interconnected filopodia that were fully stretched and adhered to the titanium surface, highly expressed F-actin was detected, and actin fibers were thickened. Under the scanning electron microscope, the cells on the smooth titanium surface were not fully adhered and stretched, and those on the TiN/Ag-coated titanium surface or the sandblasted and acid-etched titanium surface exhibited better adhesion and extension. Findings from the cell counting kit-8 showed that after 72 hours of inoculation,the cells on the smooth titanium surface grew well,with cytotoxicity level 1.In addition,Staphylococus aureus grew well on the smooth titanium surface under the confocal laser scanning microscope,while a large amount of Staphylococus aureus died on the TiN/Ag-coated titanium surface or on the sandblasted and acid-etched titanium surface. These findings indicated that TiN/Ag coating has good biocompatibility and antibacterial properties.

Objective:To study the optimal nutritional support strategy in critical patients with acute ischemic stroke.Methods:50 critical patients with acute ischemic stroke were divided into two groups.One group were started with enteral nutrition alone within 24h after admission,while the other group received sequential parenteral and enteral nutrition.The incidence of pulmonary infection,gastric retention,upper gastrointestinal bleeding,hypoglycemia and hyperglycemia were compared between the two groups.Another endpoint was discharge from ICU at day 20,and it was compared between the two groups using Kaplan-Meier methods.Results:There was no difference between the two groups in the rate of hypoglycemia or hyperglycemia (P ＞ 0.05).The incidences of pulmonary infection,gastric retention,upper gastrointestinal bleeding and diarrhea were lower in the PN+EN group than EN group (P ＜ 0.01).The length of ICU stay was also shorter in the PN+EN group.Conclusion:The strategy of sequential parenteral and enteral nutrition decreased the complication rate related to nutrition support,such as pulmonary infection,gastric retention,upper gastrointestinal bleeding.Also,it shortened the ICU stay in critical patients with acute ischemic stroke.