Chinese higher medical education has undergone a series of education reform project .College excellent courses construction is an important component of the project .To enable medical students to master the basic surgi-cal theory and skills, and to develop high -quality, high -level medical personnel , the surgery department in Shengjing Hospital of China Medical University has scored some achievements in constructing excellent courses . Some ideas and methods are summarized in the construction of excellent courses .

Animals ; Cytokines ; metabolism ; Humans ; Lung ; metabolism ; pathology ; Pulmonary Fibrosis ; metabolism ; pathology ; Reactive Nitrogen Species ; metabolism ; Reactive Oxygen Species ; metabolism

Background Proliferative vitreoretinopathy(PVR) is a tissue repair prevention and treatment of PVR in clinic.Natural delayed release microballoons are therefore becoming a hot spot for its easy manipulation,large lading dose and long acting duration.Objective This study was to evaluate the effect of 5-fluorouracil natural delayed release microballoons on the prevention of PVR.Methods The lymphocytes were collected from clean pigment rabbit to prepare the 8×107/ml cell suspension with complete culture fluid.PVR models were established in 45 healthy pigment rabbits by intravitreal injection of lymphocyte suspension.The animals were randomly divided into 3 groups and 15 rabbits for each.0.1ml normal saline,10g/L or 20g/L 5-fluorouracil natural delayed release microballoons were injected into vitreous cavity respectively.PVR was graded on Fastenberg's method under the slit lamp in 1,2,4,8 weeks.The animals were sacrificed and retinas were obtained for the histopathological and ultrastructural examination in the eighth week after administration of drug.Results The numbers of eyes with different grades of PVR were significantly different among 3 groups in 1 week,2,4,8 weeks(P＜0.05).The eye numbers with PVR was significant less in 20g/L Fu group than those of 10g/L Fu group and normal saline group(P＜0.05).There was statistical difference in PVR ranking among these 3 groups in 8 weeks after injection of drug(H=46.795,P＜0.05).The morphology and ultrastructure of retinas under the light microscope and transmission electron microscope were near normal in all of the three groups.Conclusion Implantation of 5-fluorouracil natural delayed release microballoons into vitreous cavity is effective and safe in preventing PVR in experimental model,and the therapeutic effect of microballoons with 20g/L 5-Fu is better.

BACKGROUND: At present, there has been no definite experiment systemically evaluating adherent separation and density gradient centrifugation to isolate bone marrow mesenchymal stem cells (BMSCs). Whether the two methods produce different influences on BMSC induction and differentiation remains unclear.OBJECTIVE: This study was designed to verify difference of these two isolation methods in chondrogenic differentiation of BMSCs.DESIGN, TIME AND SETTING: A controlled observation was performed at the Shengjing Hospital Affiliated to China Medical University from March to September 2005. MATERIALS: Twenty Japanese big-ear rabbits, aged 2-3 months, weighing 1.2-2.0 kg, were included for this study. METHODS: BMSCs were isolated by adherent separation and density gradient centrifugation. Two groups of BMSCs were taken from the same passage and induced towards chondrogenic differentiation with transforming growth factor beta 1. MAIN OUTCOME MEASURES: Growth of BMSCs was observed under an inverted microscope to draw growth curves; Type II collagen expression was detected by immunohistochemistry. Type II collagen mRNA expression was determined by in situ hybridization. RESULTS: The growth curves demonstrated that cellular growth velocity of the two groups tended to be the same. Immunohistochemistry results showed that the efficiency of adherent separation and density gradient centrifugation for promoting chondrogenic differentiation of BMSCs was 76.1% and 77.7%, respectively, and in situ hybridization results showed that the efficiency was 70.3% and 71.0%, respectively. No significant difference in differentiation efficiency existed between the adherent separation and the density gradient centrifugation. CONCLUSION: Adherent separation and density gradient centrifugation had no different influences on BMSC growth and chondrogenic differentiation.

Objective To observe the immunologic function of critically ill newborn and the relative function of nuclear factor kappa B(NF-?B).Methods The critically ill group contained 50 cases,and 25 cases from healthy newborns were used as control group.Blood samples were collected in each case,levels of cytokine interleukin(IL)-4,interferon(IFN)-?,tumor necrosis factor(TNF)-? and NF-?B were detected.Result Compared with control group,NF-?B of the critically ill newborn activated and the cytokine were disorder,and IL-4 and TNF-? increased,but IFN-? decreased.Conclusions Critically ill newborn exist immune functional disorder.Furthermore,NF-?B activation may be involved in the process in infants with critically illness.

The preparation method, serum stability, efficiency of cellular uptake and apoptosis induction of the cell penetrating peptide TAT and cleavable PEG co-modified liposomes loaded with paclitaxel (C-TAT-Lipo) were investigated. The best preparation procedure was performed by orthogonal test based on single factor screening method. First, the paclitaxel (PTX)-loaded liposomes were prepared by filming-rehydration method, evaluated with entrapment efficiency and polydispersity index. The morphology of C-TAT-Lipo was characterized by transmission electron microscopy. Turbidity variations were monitored in the presence of fetal bovine serum (FBS) to evaluate the serum stability of the liposomes developed here. Next, the efficiency of cellular uptake of different Rho-PE-labeled liposomes on B16F1 cells in vitro was evaluated by confocal laser scanning microscopy (CLSM) and flow cytometry. The quantitative analysis of apoptosis induced by different PTX-loaded liposomes was performed by Annexin V-FITC/PI double staining. The optimal formulation was as follows: Chol : lipid: 1 : 8 (molar ratio); drug : lipid: 1 : 40 (mass ratio); lipid concentration: 3 mmol x L(-1); temperature of hydration: 25 degrees C. The mean size and polydispersity index of C-TAT-Lipo were about (97.97 +/- 3.68) nm and 0.196 +/- 0.037, the zeta potential was (-0.89 +/- 0.45) mV, the entrapment efficiency of paclitaxel was (90.16 +/- 1.53)%. The particle sizes did not exhibit significant variations in 50% FBS over 24 h at 37 degrees C. The efficiency of cellular uptake of the C-TAT-Lipo increased 1.40 fold following the cleavage of PEG. Apoptosis analysis showed 59.3% increase of the apoptosis and necrosis profile of C-TAT-Lipo after the detachment of PEG shells, which was markedly higher than that of N-TAT-LP with or without glutathione and SL, respectively. The results indicate that the C-TAT-Lipo is successfully prepared by filming-rehydration method and shows significant antitumor activities.

Objective To evaluate the character of the collagen-chitosan-chondroitin sulfate scaffold seeded with rat adipose tissue-derived stromal cells. Methods A dipose tissue were harvested from 6 weeks old Wistar rats and the stromal cells were harvested by type Ⅰ collagenase and then cultured in vitro. Type Ⅰ collagen was fully mixed with chitosan, freeze-dried and cross-linked with chondroitin sulfate, then freeze-dried again and sterilized by ethylene oxide. The pore diameter, water content, porosity of the scaffold were tested. The adipose tissue-derived stromal cells were digested, seeded into the plates, scaffold, and cen-trifuged into pellet, and then induced into cartilage. MTT detection for cell proliferation was done. After 3 weeks, the cell morphology, and cell proliferation and adhesion were observed, and chondrngenic differenti-ation was also analyzed. Results The pore diameter, water content, porosity tested for the scaffold showed an appropriate form. Cell proliferation showed faster in the scaffold and pellet culture system after 5 day, there was still cell proliferation in the scaffold system after 14 days but no obvious changes in the pellet cul-ture system; ceils on the scaffold proliferated densely showed by histological staining, but there was a scaf-fold structure residues in the inner layer. The finding of type Ⅱ immunohistochemistry stain showed that cells express strong positive for type Ⅱ collagen in the scaffold and pellet culture system whereas it was weakly positive in the plate culture system; the specific mRNA for cartilage, type Ⅱ collagen, aggrecan and SOX-9 were expressed in all three systems showed by RT-PCR, but type X collagen was expressed continu-ously in the plate culture system and expressed after 21 days in the pellet culture system, whereas it was not detected in the collagen-chitosan-chondroitin sulfate scaffold system. Conclusion The parameters of the collagen-chitosan-chondroitin sulfate scaffold were suitable in our study. The results suggested that it can promote the adipose tissue-derived stromal cells proliferation and chondrogenic differentiation better than the plate and pellet culture systems and maintain the phenotype of chondrocytes well; it is the optimal choice for cartilage tissue engineering in the future.

BACKGROUND: The implanted cartilage calls can synthesize cartilage matrix as cartilage in cartilage tissue enginsedng, and the density of implanted cells is the key point.OBJECTIVE: To evaluate the effect of cell seeding concentration on the chondrogenic differentiation of the adipose dadved sromal cells (ADSCs).DESIGN, TIME AND SETTING: The in vitro cellular-scaffold observation was performed at the cytobiological laboratory of China Medical University from November 2007 to July 2008.MATERIALS: Six male SD rata with clean grade were supplied by the Experimental Animal Center of China Medical University.METHODS: Totally 5 g/L type ; collagen solution and 20 g/L chitosan was mixed in a mould with volume ratio of 7:3, after lyophillization, it was cut into pieces with 5 mm ～ 5 mm x 2 mm, followed by crosslinking with ethanol contained of 2% chondroitic acid at room temperature. After washing with double distilled water and freeze drying, the chitosan-collagen-chondroitin sulfate copolymar matrices scaffolds were harvested. ADSCs isolated from rat inguinal fat pads were digested with collagenase and trypsase. The prepared scaffolds were randomly divided into 3 groups, and the third passage cells with density of 2×10 9/L,2×10 109/L, and 2×10 11/L were seeded into chitosan-coflagen-chondroitin sulfate scaffolds, and cultured in chondrogenic medium for 3 weeks.MAIN OUTCOME MEASURES: The expression of cartilage specificity gene was detected by hematoxylin-eosin staining, type Ⅱ collagen immunohistochemical staining and RT-PCR.RESULTS: Hematoxylin-eosin staining showed that after 3 weeks of culture, the cell proliferated and differentiated well, especially in 2x101～/L group, more extrocelluer matrices were produced and cartilage lacuna-structure could be seen. The type Ⅱ collagen was positive expressed in each group, which showed a gradually increasing tendency with the cell seeding concentration increasing. RT-PCR showed that the expression of proteoglycen and type Ⅱ collagen mRNA were slowly increased. However the expression of Ⅹ collagen mRNA was decreased with increasing cell seeding concentration.CONCLUSION: The chitosan-collagen-chondroitin sulfate copolymer matrices can provide an appropdate environment for the generation of cartilage-like tissues and high call seeding concentration of 2×1010/L facilitate ADSCs to differentiate into cartilage.

Adult ; Female ; Humans ; Male ; Medical Staff, Hospital ; statistics & numerical data ; Middle Aged ; Nursing Staff, Hospital ; statistics & numerical data ; Sports ; statistics & numerical data

The preparation method, serum stability, efficiency of cellular uptake and apoptosis induction of the cell penetrating peptide TAT and cleavable PEG co-modified liposomes loaded with paclitaxel (C-TAT-Lipo) were investigated. The best preparation procedure was performed by orthogonal test based on single factor screening method. First, the paclitaxel (PTX)-loaded liposomes were prepared by filming-rehydration method, evaluated with entrapment efficiency and polydispersity index. The morphology of C-TAT-Lipo was characterized by transmission electron microscopy. Turbidity variations were monitored in the presence of fetal bovine serum (FBS) to evaluate the serum stability of the liposomes developed here. Next, the efficiency of cellular uptake of different Rho-PE-labeled liposomes on B16F1 cells in vitro was evaluated by confocal laser scanning microscopy (CLSM) and flow cytometry. The quantitative analysis of apoptosis induced by different PTX-loaded liposomes was performed by Annexin V-FITC/PI double staining. The optimal formulation was as follows: Chol : lipid: 1 : 8 (molar ratio); drug : lipid: 1 : 40 (mass ratio); lipid concentration: 3 mmol x L(-1); temperature of hydration: 25 degrees C. The mean size and polydispersity index of C-TAT-Lipo were about (97.97 +/- 3.68) nm and 0.196 +/- 0.037, the zeta potential was (-0.89 +/- 0.45) mV, the entrapment efficiency of paclitaxel was (90.16 +/- 1.53)%. The particle sizes did not exhibit significant variations in 50% FBS over 24 h at 37 degrees C. The efficiency of cellular uptake of the C-TAT-Lipo increased 1.40 fold following the cleavage of PEG. Apoptosis analysis showed 59.3% increase of the apoptosis and necrosis profile of C-TAT-Lipo after the detachment of PEG shells, which was markedly higher than that of N-TAT-LP with or without glutathione and SL, respectively. The results indicate that the C-TAT-Lipo is successfully prepared by filming-rehydration method and shows significant antitumor activities.
Animals ; Annexin A5 ; Apoptosis ; Cell Line, Tumor ; Cell-Penetrating Peptides ; pharmacology ; Fluorescein-5-isothiocyanate ; analogs & derivatives ; Liposomes ; chemistry ; Melanoma, Experimental ; Mice ; Microscopy, Confocal ; Paclitaxel ; pharmacology ; Particle Size ; Polyethylene Glycols ; chemistry