BACKGROUND: Silk, a natural product, has been paid wide attention in medical field owing to a fact that silk's mechanical property and biocompatibility are superior to traditional artificially synthesized degradable polymers. OBJECTIVE: To observe the effects of silk on the adsorption as well as cell morphology and function of 3T3-L1 preadipocytes. METHODS: Raw silk and trypsin-digested silk were ad Ubitum twisted into reticular fibrous cord with three-dimensional structure: mesh size 70-200 μm, thickness 200-300 μm, and porosity 20%. The three-dimensional scaffolds made of digested silk were placed in the 24-well plate. 3T3-L1 preadipocytes suspension (6×10~(10)/L) was added to the culture plate, 3 drops comprising 1 × 10~7 cells per well. Following 4-hour incubation in the air, when cells fully attached to the scaffold, they were thoroughly soaked with medium, which was renewed every other 2-3 days for a total of 1-4 weeks.RESULTS AND CONCLUSION: Inverted microscope results showed that silk scaffold/3T3-L1 preadipocytes compounds presented with slender prominences that stretched out and migrated ahead to gradually connect together through a head-to-end fusion fashion and enter into meshes. Scanning electron microscope results demonstrated that in the silk scaffold/3T3-L1 preadipocytes compounds, ceils tightly attached to silk scaffold, appropriately spread out, and secreted matrix. Silk shows better absorption for 3T3-L1 preadipocytes and maintains the normal morphology and functions of 3T3-L1 preadipocytes.

Objective To study induction of myocardial fibrosis by high-cholesterol diet and its mechanism in rat. Methods Nineteen Wister rats aged 8 weeks were randomly divided into control group, high-cholesterol diet group and spironolactone treatment group. Collagen in left ventricular tissues was determined by oxyproline assay, contents of angiotensin Ⅱ(Ang Ⅱ) and aldosterone (Ald) and in left ventricular tissue their plasm concentration were determined by radioimmunoassay. Serum nitrite level was measured by Griess assay. Mitogen activated protein kinase (MAPK) was assessed by radioisotope labeled assay. Results The contents of collagen, cardiac Ald and plasma Ang Ⅱ in the myocardium of the left ventricle were increased 1.2, 1.1 and 3.0 fold, respectively, while serum level of nitrite (NO - 2) was significantly decreased in high-cholesterol diet group compared with control group. The contents of myocardial collagen and Ald, and plasma Ang Ⅱ and the activities of MAPK were decreased in high-cholesterol diet group treated with spironolactone for 8 weeks (P

Background Sulforaphane (SFN) is an effective chemopreventive agent and can regulate the biological molecular mechanisms to inhibit the overgrowth of cells.Autophagy is a biological process of maintaining cellular internal environment.Understanding the affection of SFN to biological behavior of human lens epithelial cells (LECs) and the association of SFN with autophagy is helpful for the prevention and target treatment of posterior capsule opacification (PCO).Objective This study was to investigate the eradication effeccts of SFN on residual lens cell population in vitro posterior capsule opacification (PCO) model and evaluate the mechanism of SFN-induced cell death.Methods In vitro human capsular bag models were generated from fresh donor eyes by phacoemulsification and were cultured in EMEM containing 2％ fetal bovine serum (FBS).Different concentrations of SFN (0,1,10 and 100 μ mol) were added in the medium for 30 days respectively according to grouping,and the growth of LECs was observed by optical microscope and immunofluorescence technique.FHL124,a human LEC line,was cultured with EMEM containing 5％ FBS and divided into 0,1,10,30 and 100 μmol SFN groups.Lactate dehydrogenase (LDH) release rate in the medium was detected to evaluate cell damage/death.The migration of the cells on capsular bags was assessed by scratch test.The ultrastructure and number of autophagosomes were examined under the transmission electron microscope.The expression of LC3 in the cells were detected using Western blot in the presence or absence of autophagy inhibitors.Results The cell coverage rates on the capsular bags were significantly lower in the 10 and 100 μ mol/L SFN groups than those in the 0 and 1 μmol/L SFN groups,with a statistically significant difference among the groups (F =48.57,P ＜ 0.01).Immunofluorescence showed that the density of F-actin-and Vimentin-positive cells was evidently decreased in the 10 and 100 μmol/L SFN groups compared with 0 and 1 μ mol/L SFN groups.The releasing levels of LDH (absorbancy) were 0.19± 0.03,0.39±0.06,0.56±0.07,0.68±0.08 and 0.89±0.09 in the 0,1,10,30 and 100 μ mol/L SFN groups,respectively,and the releasing level of LDH was gradually increased in the 10 and 100 μ mol/L SFN groups in comparison with the 1 μmol/LSFN group (all at P＜0.01).With the increase of SFN concentration,the reduction rate of scratched area decreased with the increase of SFN concentration,and the decrease of scratch area was significantly lower than that of adjacent low mass concentration group and the differences were statistically significant (P＜0.05).The relative expressions of LC3-Ⅱ protein were 0.423±0.003,14.543±0.024,0.668±0.024 and 0.576±0.056 in the blank control group,SFN group,SFN + 3-MA group and 3-MA group,respectively,and the relative expressions of LC3-Ⅱ protein were significantly lower in the SFN+3-MA group and 3-MA group than those in the SFN group (all at P＜0.01).The number of autophagosomes was 4.07±0.32,4.13±0.34,9.21 ±0.53 and 21.02± 1.34 in the blank control group,and 1,10,100 μmol/L SFN groups,and the number of autophagosomes in the 10 and 100 μ mol/L SFN groups was significantly higher than that in the blank control group and 1 μmol/L SFN group (all at P＜0.01).Conclusions SFN mediates LECs death by promoting autophagy in ex vivo capsular bags,and SFN may be a novel agent of potential chemopreventive and target treatment for PCO.

Objective To investigate the changes of serum inflammatory mediators in septic rats. Methods Thirty male SD rats were randomly divided into sham operation group(10 rats)and sepsis group (20 rats). The model of sepsis was made by cecal ligation and puncture. Carotid artery blood samples were taken from the sham operation group,and from sepsis group at 0,24,48,72 h after model establishment. Enzyme linked immunosorbent assay(ELISA)was applied to detect the content of serum tumor necrosis factor alpha(TNF α), interleukin 1(IL-1),interleukin element 6(IL-6). Results Serum TNF-α content in the sham operation group was(9. 27 ± 3. 12)ng/ L,and(9. 26 ± 8. 01),(32. 01 ± 4. 52),(55. 22 ± 7. 61),(83. 31 ± 8. 57)ng/ L respectively at 0,24,48,72 h after model establishment in sepsis group. There were significant difference between sham operation group and sepsis group at different time points(P < 0. 01). Serum IL-1in sham operation group was(8. 93 ± 1. 26)ng/ L,lower than that in sepsis group model at 0,24,48,72 h after model establishment ((20. 01 ± 3. 51),(25. 51 ± 2. 79),(59. 67 ± 3. 26),(87. 86 ± 11. 51)ng/ L respectively),and the differences were significant(P < 0. 01). The serum IL-6 level in sepsis group at 0,24,48,72 h after model establishment were(11. 52 ± 2. 32),(31. 59 ± 12. 12),(57. 27 ± 13. 53),(71. 59 ± 12. 67)ng/ L,respectively,different from that of sham operation group((12. 36 ± 3. 25)ng/ L,(P < 0. 01)). Conclusion The serum transmitter levels in septic rats significantly increase,which suggest that a lot of inflammatory mediators are released,and it may be one of the factors for the occurrence of sepsis.

Recently, along with the harmful factors in air in-creasing, the incidence of respiratory system diseases has been gradually rising. Therefore, more studies are focused on impro-ving the prevention and treatment on respiratory system diseases. Current researches have shown that microRNAs ( miRNA ) con-tribute to the prevention and treatment on respiratory system dis-eases, including virus infectious respiratory disease, pulmonary fibrosis, asthma, COPD, pulmonary tuberculosis and lung canc-er. The up-regulation and down-regulation of microRNAs could effectively regulate the biological process of these diseases. The review analyzes the recent years′study results of respiratory sys-tem diseases and microRNAs in depth, and discuss the role of microRNA in the treatment and prevention of respiratory disea-ses. Moreover, in this review we can provide new knowledge a-bout respiratory diseases and development of more potent drug targets for the treatment and prevention of respiratory diseases.

BACKGROUND:Preadipocyte is a kind of specific precursor cells with the potential of proliferation and differentiation into adipocytes.If factor or drug with strong promotion of preadipocyte proliferation and differentiation is obtained and used in adipose tissue transplantation,which could elevate transplantation outcome of adipose tissues.OBJECTIVE:To find an optimal culture environment and to accumulate culture experience of in vitro culture 3T3-L1 preadipocytes,and to observe the effect of insulin-like growth factor(IGF)-1 on the proliferation and differentiation of 3T3-L1 preadipocytes,and to explore the optimal concentration of IGF-1.DESIGN,TIME AND SETTING:The cell control observation was performed at the Science Experiment Center,Liaoning University of Traditional Chinese Medicine from October 2007 to February 2008.MATERIALS:3T3-L1 preadipocytes were purchased from Shanghai Institutes For Biological Sciences.IGF-1 was bought from Biological,USA.METHODS:According to in vitro culture direction of 3T3-L1 preadipocytes,3T3-L1 preadipocytes were divided into 6 groups.3T3-L1 preadipocytes in the blank control group were inoculated in high-glucose DMEM supplemented with 0.1 volume fraction of fetal bovine serum.3T3-L1 preadipocytes in each experimental groups were respectively treated with high-glucose DMEM containing 5,10,20,50,100 ?g/L IGF-1 and 0.1 volume fraction of fetal bovine serum.MAIN OUTCOME MEASURES:Growth,proliferation and differentiation of 3T3-L1 preadipocytes in each experimental groups under the inverted microscope,and photographed.When cells were confluence,cell cycles were detected by flow cytometry.Growth rate of 3T3-L1 preadipocytes was measured by MTT essay.Fat growth rate in 3T3-L1 preadipocytes were examined by using oil red O stained-extracted assay.RESULTS:Inverted microscope showed that 3T3-L1 preadipocytes before differentiation were fusiform shape,with little fat in their endochylemas.3T3-L1 preadipocytes after differentiation were spherical shape,with much fat in their endochylemas.Lipid droplet gathered in the perinuclear space,and was stained orange by oil red O stained-extracted assay,forming ring-shape.Results from MTT essay and oil red O stained-extracted assay demonstrated that growth rate of 3T3-L1 preadipocytes and fat growth rate in 3T3-L1 preadipocytes were higher in each experimental groups treated with different doses of IGF-1 compared with the blank control group(P

0.05).10% FBS induced -TdR incorporation increased by 87.5%(vs control,P

Objective To establish a method of adsorption and thermal desorption-GC-MS and to study the component of organic pollutants in indoor air in newly decorated houses. Methods Used Tenax TA as an adsorbent, the qualitative analysis of the indoor air in two typical newly decorated houses had been done with adsorption-thermal desorption GC-MS method. Results More than sixty organic compounds were detected, besides volatile organic compounds such as acetic acid, glycol, toluene and semivolatile organic compounds such as acenaphthene, anthracene, octadecane, monocyclic aromatic hydrocarbons, polycyclic aromatic hydrocarbons, fatty hydrocarbons, oxygen contained hydrocarbons and so on were detected. Conclusion This method can be used for the qualitative analysis of pollutants in indoor air. The organic compounds detected in the air of newly decorated houses are complex of different components.

AIM:To investigate whether hypoxic preconditioning(HPC)protects cardiomyoblast H9c2 cells against oxidative injury,and to discuss whether calreticulin(CRT)contribute to this protection through p38 MAPK signaling pathway.METHODS:Cardiomyoblast H9c2 cells were randomly divided into eight groups as follows:hydrogen peroxide stress(H2O2);brief hypoxic exposure of 20 min to simulate hypoxic preconditioning(HPC);20 min of hypoxic exposure followed by 24 h of normoxic reoxygenation before hydrogen peroxide stress(HPC+H2O2),SB203580(the specific inhibitors of p38 MAPK)+HPC+H2O2,antisense oligonucleotides transfection of calreticulin(AS),AS+H2O2,AS+HPC+H2O2 and control.Morphological studies,estimation of lactate dehydrogenase(LDH)leakage and flow cytometry were employed to assess the cell apoptosis and necrosis.RT-PCR and Western blotting analysis was used to detect calreticulin expression and phosphorylation of p38 MAPK.RESULTS:The results obtained are as follows:(1)HPC relieved cell injury caused by H2O2.Compared with those in H2O2 group,apoptosis rate and LDH leakage in culture medium in HPC + H2O2 group decreased 13.4% and 44.0%,respectively(P

AIM To explore the inhibitory effects of simv astatin on ROS generation and cardiac myocytes hypertrophy induced by ET-1. METHODS The study was performed with primary cultured neonatal rat cardiac myocytes. Intrace llular fluorescence signal was assayed by fluorescence convert microscope. The l evel of intracellular ROS was measured by the ROS-specific probe 2′, 7′-dich lorofluorescin diacetate (DCF-DA). The RNA content was determined by RNA-sensitive fluoresce nce probe propidium iodide (PI) and the total protein of cell was measured by th e methods of coomassie brilliant blue. The cell surface area was measured by ima ge analysis program. RESULTS ①Fluorescence intensity of intracel lular DCF-DA and myocyte hypertrophy increased by ET-1 in dose-dependent mann er. Antioxidant catalase(0 2 U?L -1 ) attenuated the ET-1-induced incre ase of fluorescence intensity of intracellular DCF-DA and myocyte hypertrophy.②Simvastatin inhibited the increase of fluorescence intensity of intracellular DCF-DA and cardiac myocyte hypertrophy induced by ET-1(10 -8 mol?L -1 )in a dose-dependent manner. CONCLUSION ET-1 increases int racellular ROS in the cultured neonatal rat cardiac myocytes. Simvastatin inhibi ted the generation of ET-1-induced ROS and cardiomyocytes hypertrophy.