Objective To study the relationship among apoptosis,NF-KB activation and uPA expres- sion in human colon carcinoma cell line HCTll6 induced by 5-fluorouracil,and to observe the effect of in- hibiting activity of NF-KB by PDTC on apoptosis as well as expression of uPA.Methods Cell apoptosis was analysed by Annexin V-FITC.Fluctuation of NF-KB and uPA was detected by semi-quantitative immuno- histochemistry.Results 5-fluorouracil could induce apoptosis and activate NF-KB.PDTC could significantly increase the apoptosis and suppress the activation of NF-KB induced by 5-fluorouracil.There was a positive correlation between the changes of uPA and NF-KB.Conclusion 5-fluorouracil could induce apoptosis,ac- tivate NF-KB and up-regulate expression of uPA of HCT116 cells.The mechanism of enhanced apoptosis by PDTC may be related to suppressing activation of NF-?B and down-regulating expression of uPA.

This study was aimed to establish the quality standard of Pyrethri Tatsienenis Flos. The medical material was identified by the microscopy and the thin layer chromatography ( TLC ) methods . The moisture , total ash , acid-insoluble ash and alcohol-soluble extract were determined according to procedures recorded in the Chi-nese Pharmacopoeia (2010 edition). The content of luteolin was determined by the HPLC method. The results showed a strong characteristic microscopic of Pyrethri Tatsienenis Flos , and its TLC identification had a good resolution with clear spots . The mass fractions of luteolin was 0 . 036%~0 . 104% ( average of 0 . 078%) , moisture was 9 . 32%~15 . 82% ( average of 13 . 11%) , total ash was 6 . 65%~8 . 29% ( average of 7 . 45%) , acid-insoluble ash was 0 . 23%~0 . 59% ( average of 0 . 42%) , and the extraction was 21 . 42%~30 . 15% ( average of 24 . 86%) . It was concluded that this established standard was simple to operate with good stability and reproducibility , which can be used for quality evaluation of Pyrethri Tatsienenis Flos .

Objective To evaluate the early clinical efficaoy of hybrid procedure for infants less than six months old with ventricular septal defect and coarctation of aorta.Methods From January 2010 to July 2011,20 patients with ventricular septal defect and coarctation of aorta received hybrid procedure in our center.The body weight was (4.5 ± 1.6) kg ( ranged from 1.9 kg to 6.5 kg) and the age was ( 56 ± 45 ) days ( ranged from 18 days to 6 months).The pressure gradient of the coarctation of the aorta ranged from 30 mm Hg to 56 mm Hg,5 patients of them were diagnosed as hypoplasty of aortic arch.The size of the ventricular septal defect ranged from 8 mm to 16 mm.Results The mortality was zero in all the 20 cases during the surgery,and the mobidity was 20％ (4/20).The complications were pneumonia in 2 cases,infective endocarditis in 1 case and pneumothorax in 1 case.The diameter of coarctation of the aorta ranged from 1.5 mm to 3.4 mm,and the size of the balloon ranged from 4 mm to 12 mm.The pressure gradient of the coarctation of the aorta decreased to 0 to 27 mm Hg.The bypass time ranged from 40 minutes to 87 minutes,and the crossclamp time of the aorta ranged from 20 minutes to 41minutes.The atrial septal defects were repaired and the patent ductuses were ligated during the surgery without leaving the sternum open.The total operation time was (4.0 ± 0.7 ) hours ( ranging from 3.0 hours to 5.2 hours).The mean ventilation time was (2.2 ± 1.4) days and mean ICU stay time was (5 ± 3 ) days.All the patients were followed up for ( 10.0 ± 3.6) months without aneurysm in arch and obstruction in airway.The residual obstructive pressurc gradicnt in the aortic arch ranged from 12 mm Hg to 35 mm Hg and 2 patients received reintervention.One patient received re-balloon dilation and the other received surgery.The cardiac function reached NYHA Ⅰ - Ⅱ in all eases.Conclusion The early outcome of the hybrid procedure (balloon dilation of the coartation of the aorta and surgical repair of ventricular septal defect) for infants with ventricular septal defect and coarctation of aorta was satisfying,which could avoid from circulatory arrest.It is a relatively safe procedure which could be the optional method for one-stage surgical repair.

BACKGROUND:Bone morphogenetic protein-2 (BMP-2) is a key to bone formation and repair.However,it has some disadvantages such as easy to lose and degrade and difficult to sustain continuous effect.OBJECTIVE:To study the preparation and properties of silk fibroin/chitosan/nano-hydroxyapatite (SF/CS/nHA) scaffold loading BMP-2.METHODS:After silk degumming,dissolution and purification,2％ SF solution was obtained.BMP-2 was dissolved in 2％ CS solution,and then fully mixed with equal volume of SF solution and proper amount of nHA.At last,the SF/CS/nHA scaffold loading BMP-2 was prepared using freeze-drying method as experimental group.The SF/CS/nHA scaffold was soaked in the BMP-2 solution as control group.The scaffold porosity was measured by Archimedes method,the surface morphology of the scaffold was observed by scanning electron microscope,the compressive strength was measured by universal testing machine.Scaffolds in the two groups were soaked in PBS,and the release of BMP-2 was measured by ELISA method at different time points.RESULTS AND CONCLUSION:(1) The scaffolds in the two groups had irregular porous structure,interconnected pores and uneven pore wall.There was no significant difference between the two groups in mean pore diameter,porosity and maximum compressive strength.(2) On the 1st day,the release rate of BMP-2 was 4.63％ in the experimental group,and the release curve increased slowly.After 28 days,the release curve of BMP-2 was transferred to the plateau stage.But in the control group,the release rate of BMP-2 on the 1stday was 58.84％,and it was a significant initial burst release.The release curve increased rapidly,and was transferred to the platform stage on the 10th day.The release rate of BMP-2 release was significantly different between the two groups at days 1,2,4,10 (P ＜ 0.05).These results show that the SF/CS/nHA scaffold loading BMP-2 could sustainably and slowly release BMP-2.

Objective To investigate the reaction kinetics between the stone-dissolving solution and the urinary stone in a model simulating the condition of a kidney for further clinical administration. Methods An artificial upper urinary tract was made by silica gel. Lactic acid prepared in the preliminary study was used to react with artificial stone in the model. The concentration of reaction product in the effluent was measured to identify the reaction velocity. Relationships between the efficiency and dissolution rate or stone surface was investigated. Results The highest utilization rate of dissolution was 100 ~ 150 mL/h. Dissolution efficiency is in positive rela-tion with stone surface. The efficiency correlates with the stone surface and infusion speed in the range of 50~400 mL/h. Conclusions Before dissolution treatment ,the stone should be shattered as deeply as possible to in-crease the surface of reaction. If possible ,the irrigating speed should be as high as possible to eliminate the stone sooner.

With the rapid development of the biopharmaceutical industry in China, the laboratory animal sector has entered a stage of rapid growth, and the construction of facility operation systems has become increasingly automated and intelligent. Compared to traditional laboratory animal facilities, new facilities require a more specialized technical team for the maintenance of air supply and exhaust systems, air conditioning, automated control, and the entire barrier system. The Lingang Laboratory’s animal facilities accommodate both large and small animals for feeding and experimental purposes. The facility management team has summarized daily maintenance experiences and explored various operational and maintenance modes based on the characteristics of laboratory operation. After analyzing the advantages and disadvantages of three common modes, this paper provides new ideas for the management of these laboratory animal facilities, and offers guidance for peers in choosing the most appropriate professional maintenance mode.

Animals ; Child ; Child, Preschool ; Dermatitis ; epidemiology ; parasitology ; Disease Outbreaks ; Female ; Humans ; Male ; Mites ; Schools, Nursery

OBJECTIVETo investigate the effects and regulatory mechanism of ILK on angiogenesis in hypertrophic scar.

METHODSThe human scar microvascular endothelial cells (HSMECs) were isolated from 6 patients' hypertrophic scar in vitro. The HSMECs with good condition in 2nd to 4th generation were selected as experimental objectives. (1) HSMECs were divided into the blank control group (treated with routine culture), negative control group (treated with only Lipofectamine 2000), LY294002 group (incubated with 50 nmol/L LY294002), ILK siRNA group (incubated with 20 nmol/L ILK siRNA). RT-PCR and Western Blot were used to detect the expression of ILK mRNA and its protein after transfecion for 48 h. (2) The digested HSMECs of four groups were resuspended with DMEM without serum and then seeded onto the upper compartment of transwell insert which contained complete medium in its lower compartment. The cell migration experiment was stopped in 10 h and then the migrated cells were counted to analyze the effects of different interventions on the migration ability of HSMECs. (3) The thawed ECMatrix was put into each well of pre-colled 48-well tissue culture plate, and then the plate was put into the incubator at 37 degrees C to make it to become gel. The HSMECs of four groups were seeded onto the surface of the ECMatrix gel and were put into incubator. Eight random view-fields per well should be valued by the sheet of pattern recognition about angiogenesis after 8 hours to evaluate the ability of angiogenesis in vitro between four groups.

RESULTS(1) The expression of ILK mRNA (ILK mRNA = 0.829 +/- 0.109, t = 13.151, P = 0.006) and protein (ILK protein = 0.096 +/- 0.049, t = 36.656, P = 0.000) were both inhibited obviously in ILK siRNA group compared with the blank control group (ILK mRNA = 0.829 +/- 0.109, ILK protein = 1). And, the expression of ILK in LY294002 group was slightly lower than that of black control group, but there was no statistical difference. (2) The number of migrated cells in ILK siRNA group (88.111 +/- 3.079) and LY294002 group (138. 667 +/- 2.404) were respectively lower than that in blank control group (322.333 +/- 3.712, P < 0. 05) in 10th hour. (3) Compared to blank control group (4.333 +/- 0.191), the ability of angiogenesis in vitro decreased significantly ILK siRNA group (2.625 +/- 0.125) and LY294002 group (3.125 +/- 0.250), in which, the vascular network structures were not formed perfectly in 8th hour (P < 0.05).

CONCLUSIONSThe ability of HSMECs' migration and angiogenesis in vitro are inhibited significantly when the expression of ILK is down-regulated. It reveals that ILK may play an role in the regulation of scar angiogenesis.

Cell Movement ; Cell Proliferation ; Chromones ; pharmacology ; Cicatrix, Hypertrophic ; enzymology ; pathology ; Endothelial Cells ; cytology ; drug effects ; Humans ; Lipids ; pharmacology ; Morpholines ; pharmacology ; Neovascularization, Pathologic ; etiology ; pathology ; Protein-Serine-Threonine Kinases ; genetics ; physiology ; RNA, Messenger ; analysis ; RNA, Small Interfering ; metabolism

By reverse transcription-polymerase chain reaction (RT-PCR), an open reading frame of pathogenesis-related protein 1 (PR1) was isolated from Panax notoginseng and named as PnPR1. Molecular and bioinformatic analyses of PnPR1 revealed that an open reading frame of 501 bp was predicted to encode a 166-amino acid protein with a deduced molecular mass of 18.1 kD. Homology analysis showed that the deduced amino acid sequence of PR1 protein of Panax notoginseng had a high similarity with other higher plants had the same conservative structure domain of cysteine-rich secretory protein (CAP). The recombinant expressed plasmid pET28a(+)-PnPR1 was expressed in Escherichia coli BL21. The expression conditions were optimized by induction at different times, different temperatures, different IPTG concentrations and different giving times. The optimum expression condition was 0.4 mmol.L-1 IPTG at 28 degrees C for 20 h. The successful expression of PnPR1 provides some basis for protein purification and preparation of the monoclonal antibody.
Amino Acid Sequence ; Cloning, Molecular ; Escherichia coli ; metabolism ; Molecular Weight ; Open Reading Frames ; genetics ; Panax notoginseng ; chemistry ; Phylogeny ; Plant Proteins ; genetics ; metabolism ; Plants, Medicinal ; chemistry ; Reverse Transcriptase Polymerase Chain Reaction ; Sequence Alignment

OBJECTIVEThe purpose was to compare two different ways of culturing hair follicle bulge stem cell: The defined keratinocyte-serum free medium (DK-SFM) method and the 3T3 feeder cell method.

METHODSThe morphological features of cultured bulge stem cells were investigated by inverted phase control microscopy. Immunostaining of stem cell marker cluster of differentiation 34 (CD34) and epithelial cell marker cytokeratin 19 (CK19) were performed to identify the bulge stem cell. The stemness of bulge stem cells was evaluated by colony forming efficiency (CFE) and proportion of CD34 positive cells by flow cytometry.

RESULTSHair follicle bulge stem cells could be successfully cultivated in vitro using two methods. They were both positive for CK19 and CD34. The colony forming efficiency of hair follicle stem cell cultured in DK-SFM and the 3T3 feeder cell was 69.4% and 62.2%, respectively. There was no significant difference in colony forming efficiency between these two methods (P > 0.05), while the CD34 positive cells proportion was higher in DK-SFM as 72.3% than the other as 34.7% (P < 0.05).

CONCLUSIONTwo methods are applicable to culture bulge stem cells in vitro. The 3T3 feeder cell method is complicated and can propagate a lot bulge stem cells from hair follicle, while the DK-SFM method is easier to get pure bulge stem cell.

Cell Culture Techniques ; Cell Differentiation ; Cells, Cultured ; Epithelial Cells ; Hair Follicle ; Keratinocytes ; Stem Cells