Osteogenic Response of MC3T3-E1 and Raw264.7 in the 3DEncapsulated Co-Culture Environment

BACKGROUND@#Three-dimensional (3D) in vitro cultures recapitulate the physiological microenvironment and exhibit high concordance with in vivo conditions. Improving co-culture models with different kind of cell types cultured on a 3D scaffold can closely mimic the in vivo environment. In this study, we examined the osteogenic response of pre-osteoblast MC3T3-E1 cells and Raw264.7 mouse monocytes in a 3D-encapsulated co-culture environment composed of the Cellrix®3D culture system, which provides a physiologically relevant environment. @*METHODS@#The Cellrix® 3D Bio-Gel scaffolds were used to individually culture or co-culture two type cells in 3D microenvironment. Under 3D culture conditions, osteoblastic behavior was evaluated with an ALP assay and staining. ACP assay and TRAP staining were used as osteoclastic behavior indicator. @*RESULTS@#Treatment with osteoblastic induction factors (?3F) and RANKL had on positively effect on alkaline phosphatase activity but significantly inhibited to acid phosphatase activity during osteoclastic differentiation in 3D coculture. Interestingly, alkaline phosphatase activity or acid phosphatase activity in 3D co-culture was stimulated with opposite differentiation factors at an early stage of differentiation. We guess that these effects may be related to RANK– RANKL signaling, which is important in osteoblast regulation of osteoclasts. @*CONCLUSION@#In this study, the osteogenic response of 3D encapsulated pre-osteoblast MC3T3-E1 cells and mouse monocyte Raw264.7 cells was successfully demonstrated. Our 3D culture conditions will be able to provide a foundation for developing a high-throughput in vitro bone model to study the effects of various drugs and other agents on molecular pathways.

Osteogenic Response of MC3T3-E1 and Raw264.7 in the 3DEncapsulated Co-Culture Environment

BACKGROUND@#Three-dimensional (3D) in vitro cultures recapitulate the physiological microenvironment and exhibit high concordance with in vivo conditions. Improving co-culture models with different kind of cell types cultured on a 3D scaffold can closely mimic the in vivo environment. In this study, we examined the osteogenic response of pre-osteoblast MC3T3-E1 cells and Raw264.7 mouse monocytes in a 3D-encapsulated co-culture environment composed of the Cellrix®3D culture system, which provides a physiologically relevant environment. @*METHODS@#The Cellrix® 3D Bio-Gel scaffolds were used to individually culture or co-culture two type cells in 3D microenvironment. Under 3D culture conditions, osteoblastic behavior was evaluated with an ALP assay and staining. ACP assay and TRAP staining were used as osteoclastic behavior indicator. @*RESULTS@#Treatment with osteoblastic induction factors (?3F) and RANKL had on positively effect on alkaline phosphatase activity but significantly inhibited to acid phosphatase activity during osteoclastic differentiation in 3D coculture. Interestingly, alkaline phosphatase activity or acid phosphatase activity in 3D co-culture was stimulated with opposite differentiation factors at an early stage of differentiation. We guess that these effects may be related to RANK– RANKL signaling, which is important in osteoblast regulation of osteoclasts. @*CONCLUSION@#In this study, the osteogenic response of 3D encapsulated pre-osteoblast MC3T3-E1 cells and mouse monocyte Raw264.7 cells was successfully demonstrated. Our 3D culture conditions will be able to provide a foundation for developing a high-throughput in vitro bone model to study the effects of various drugs and other agents on molecular pathways.

Effect of Buyang Huanwu Tang on Axonal Regeneration of Rats After Ischemic Stroke Injury / 中国实验方剂学杂志

Objective:To investigate the effects of Buyang Huanwu Tang （BHT） on axonal regeneration and neurological rehabilitation of the rats suffering ischemic stroke （IS）. Method:A total of 180 SD rats were used to establish a middle cerebral artery infarction （MCAO） model. The animals that were successfully modeled were randomly divided into model group， BHT group （12 g·kg-1） and nimodipine group （20 mg·kg-1）， and a sham group was established， with 28 rats in each group. After seven-days intragastric administration of BHT， the animals were sacrificed. TTC staining was used to test cerebral infarction. Brain water content was measured to observe cerebral edema. Bielschowsky's silver staining and immunofluorescence were performed to observe axonal degeneration and the protein expression of neurofilament protein-200（NF-200）. Quantitative real-time polymerase chain reaction （PCR） was used to analyze the mRNA expression of repulsion oriented molecule a （RGMa）， Ras homologous enzyme （Rho）， Rho kinase （ROCK）， and collapsion response regulatory protein 2 （CRMP2）. Neurological function scores assay was used to examine neurological recovery. Result:Compared with sham group， the cerebral infarction volume and brain water content increased significantly（P<0.01）， and motor function was markablely decreased in the model group. Axonal degeneration and nerve fiber damage were obviously observed. Also， gene expression of axon growth-related protein was deviation from normal （P<0.01）. Compared with model group， the cerebral infarction rate （P<0.01）， brain water content （P<0.01） and axonal degeneration of BHT group and nimodipine group were significantly reduced. The expression of NF-200 was increased. Also， the mRNA expression of RGMa， Rho and ROCK was lower （P<0.05） while the mRNA expression of CRMP2 was higher （P<0.01）. And the neurological function was significantly improved （P<0.05）. Conclusion:BHT can promote axon regeneration after ischemic stroke injury by regulating the mRNA expression of axon growth-related protein， thereby improving nerve function.

The distribution and drug resistance of respiratory tract infection pathogens isolated from patients of pediatric CICU / 国际检验医学杂志

Objective To analyze the common respiratory tract infection pathogens distribution and their drug resistance in pedi-atric cardiac intensive care unit(CICU),so as to provide reference for clinical rational use of antibiotics.Methods 1 350 cases of sputum specimens from lower respiratory tract infection patients of pediatric CICU in the medical center between January 2011 and December 2012 were cultivated and drug susceptibilities were tested.The results were retrospectively analyzed.Results 490 patho-genic strains were isolated from 1 350 cases of sputum specimens and identified,including Gram negative bacilli 288 strains (58.78%),Gram positive coccus 140 strains(28.57%),fungi 62 strains(12.65%,mainly Candida albicans ).Gram negative bacilli was given priority to with Klebsiella pneumoniae (62 strains,12.65%),followed by Branhamella catarhalis ,Pseudomonas aerugi-nosa and Escherichia coli .The rates of extended-spectrum beta-lactamases(ESBLs)-producing strains among Escherichia coli and Klebsiella pneumoniae were 73.33% and 66.13%,respectively.Gram positive coccus was given priority to with Staphylococcus aureus (65 strains,13.27%),followed by Streptococcus pneumoniae .Methicillin-resistant Staphylococcus aureus (MRSA)accounted for 24.62%.Conclusion Staphylococcus aureus ,Streptococcus pneumoniae and Klebsiella pneumoniae are main pathogens of re-spiratory tract infection in pediatric CICU.And there is multiple drug-resistant bacteria infection.Rational applicattion of antibiot-ics according to the test results of isolation and drug susceptibility is an effective way to control the infection of critical children and reduce the emergence of resistant strains.

Orthogonal design based optimization of a mouse model of acute liver failure induced by D-galactosamine and lipopolysaccharide / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To apply an orthogonal design optimization strategy to a mouse model of acute liver failure induced by D-galactosamine (D-GalN) and lipopolysaccharide (LPS) exposure.</p><p><b>METHODS</b>A four-level orthogonal array design (L16(45)) was constructed to test factors with potential impact on successful establishment of the model (D-GalN and LPS dosages, and dilution rate of the D-GalN/LPS mixture). The mortality rate of mice within 24 hours of D-GalN/LPS administration was determined by the Kaplan-Meier method. The model outcome was verified by changes in serum alanine transferase level, liver histology, and hepatocyte apoptosis.</p><p><b>RESULTS</b>The orthogonal array identified the optimal model technique as intraperitoneal injection of a combination of D-GalN and LPS at dosages of 350 mg/kg and 30 mug/kg, respectively, and using a dilution rate of 3. The dosages tested had no effect on survival. The typical signs of liver failure appeared at 6 hrs after administration of the D-GalN/LPS combination.</p><p><b>CONCLUSION</b>The orthogonal design optimization strategy provided a procedure for establishing a mouse model of acute liver failure induced by D-GalN and LPS that showed appropriate disease outcome and survival, and which will serve to improve future experimental research of acute liver failure.</p>

THE PROTECTIVE EFFECT OF TEA POLYPHENOLS ON RENAL DAMAGE IN RATS INDUCED BY D-GALACTOSE AND ITS MECHANISM / 营养学报

Objective To investigate the effect of tea polyphenols (TP) on renal damage in rats model induced by D-galactose. Methods Rats were injected with D-galactose (150 mg/kg?d),ip for 8 w,to induce renal damage. From the 3rd week,TP (150,75,37.5 mg/kg?d),aminoguanidine (150 mg/kg) and vitamin E (150 mg/kg) were administered with D-galactose for 6 w. After treatment,fasting blood glucose and 2 h blood glucose in oral glucose tolerance test were measured. The levels of HbA1C and fructosamine in serum,the activity of aldose reductase and content of advanced glycation end products (AGEs) in plasma and in kidney tissues and the activity of SOD,GSH-Px,and the contents of MDA in kidney tissues were measured,and 24h urinary protein,blood urea nitrogen (BUN) and serum creatinine (Cr) were detected. The apoptosis of renal cells were detected by flow cytometer. Results After treatment of D-galactose for 8 w,2h glucose level in oral glucose talerance test was increased significantly,the activity of aldose reductase and the content of AGES were increased significantly in blood. The levels of AGEs and MDA in renal tissues were also enhanced significantly. However,the activities of SOD and GSH-Px decreased. Additionally,the contents of 24h urine protein,BUN,Cr and the apoptotic rate of renal cells were increased significantly. High and middle dose of TP could can decrease the activity of aldose reductase in red blood cells,and inhibit the formation of glycation products in model rats induced by D-galactose. Also,TP could enhance the antioxidative activities and decrease the contents of AGEs and MDA in renal tissues. Mesnwhile,24h urine protein,BUN and Cr and the apoptotic rate of renal cells were increased significantly. Conclusion TP can inhibit glycation reaction induced by D-galactose and then protect renal from damage caused by glycation.