AIM: A novel approach to generate reference fingerprint, named principal component analysis method(PCA), was proposed in this paper. METHODS: Chromatographic finerprint of Tongbiding Injection and multi-wavelength chromatographic fingerprint of Fructus Gardenia were served as examples. Their corresponding reference fingerprints were yielded via PCA, and by comparison with average method and median method were also performed. RESULTS: Reference fingerprint, yielded by PCA based on least-squares principle, was essentially the linear combination of original chromatographic fingerprints. It had a better capability of maintaining and summarizing the data information of original chromatographic fingerprints and optimized properties, and was very similar to one generated by average method or median method. Abnormal fingerprints, if were possibly present in population of fingerprints, would be conveniently found via PCA method, and treated by using appropriate methods to eliminate the effects on reference fingerprint yielded. CONCLUSION: PCA method can be served as an effective approach to produce the reference fingerprint.

Objective: To prepare surface with collagen, so as to increase the histocompatibility of deantigenic bovine cancellous bone. Methods: Bovine cancellous bone was degreased and macerated by H 2O 2 and decalcified with 0.3 mol/L HCl to form scaffold, surface of which was treaed with collagen, then cultured rabbit osteoblasts were seeded into the scaffold and cultured in vitro . 4, 10, and 21 days later, the cells on the composite materials were observed under sanning election microscope. Results: On day 4, the cells grew in the surface of the scaffold; on day 21, the cells grow into the pores and on all of the surace of the scaffold.Conclusion: Deantigenic bovine cancellous bone with surfaces treated by collagen is histocompatable with rabbit osteoblasts.

To establish a xenografted tumor strain of human osteosarcoma in nude mice and a homologous cell line, the osteosarcoma speciments from patients were inoculated into the subcutaneous tissue of nude mice. After the transplanted tumor had grown to about 2cm in diameter, the tumor tissue was cultured in vitro and the continuously growing cells were analyzed by morphology, histochemistry, immunohistochemical straining, cell cycling analysis ,chromosome analysis and heterotransplantation. A newly established cell line designated PL 1 was thus obtained in this laboratory in continuous culture for over 100 passages. Under light and electron microscopes, the cells assumed the main characters of malignancy.Morphological observation, histochemistry analysis and BMP immunohisto chemical straining showed that they had the features of osteosarcoma.The cell cycle analysis showed 53 2% of the cells were in G 1. This cell line could produce ALP and BMP.Chromosome analysis confirmed that they had retained a karyotype of human cancer cells and a hypotriploid feature with a mode number of around 64～66.The tumor formation rate after heterotransplantation was 100%. It had lung metastasis characters. Therefore, PL 1 cell line derived from the xenograft in nude mice has been established and maintained for over 9 months trough 100 passages, and this cell line can provide material and model for further investigation of human osteosarcoma.

In the present study, a new compound named 17-(6-cinnamamido-hexylamino-)-17-demethoxygeldanamycin (CDG) was obtained by introducing the cinnamic acid (CA) group into the 17-site of geldanamycin (GDM). The anti-cancer effects of CDG in vitro and in vivo were evaluated. MTT assay was used to examine the inhibitory effect of CDG on the proliferation of MCF-7, HepG2, H460 and SW1990 cells. Immunofluorescent staining flow cytometry combined with Annexin V-FITC/PI staining were used to detect apoptotic cells. Transwell assay was used to analyze the effect of CDG on cell invasion and migration ability. Western blotting was used to detect the expression levels of RAF-1, EGFR, AKT, CDK4 and HER-2 of MCF-7, HepG2 and H460 cells. The toxicities of CDG and GDM were evaluated in mice. Using the subcutaneously transplanted MCF-7 xenograft in nude mice, inhibitory effect was evaluated in vivo. The results showed that CDG inhibited the proliferation of cancer cells (IC50: 13.6-67.4 microg.mL-1). After exposure to CDG for 48 h, most cells presented typical morphologic changes of apoptosis such as chromatin condensation or shrunken nucleus. The rates of apoptosis of MCF-7, HepG2, H460 and SW1990 cells incubated with 10 microg.mL-1 CDG were 23.16%, 27.55%, 22.21%, 20.47%, respectively. A dose-dependent reduction of migration of four cell lines was found after exposure to CDG. The decreased levels of RAF-1, EGFR, AKT, CDK4 and HER-2 showed that CDG possessed HSP90 inhibitory effect. The result of animal toxicity test on the mice suggested that CDG had lower toxicity than GDM. Meanwhile, CDG inhibited the growth of MCF-7 xenografts of athymic mice.

Objective To investigate the effects of AGEs-RAGE on the apoptosis of GLUTag cells and explore the possiblc mechanism.Methods GLUTag cells treated with 0、100、200、300μg/ml of AGEs for 24h were examined for gene and protein expression of RAGE using RT-PCR and western blotting,respectively.GLUTag cells were randomly divided into four groups:control,200μg/ml AGEs,AGEs+siRNA-RAGE and AGEs+apocynin.The protein expression of p22phox、p47phox 、Bcl-2、Bax in the cells were detected with western blotting.The reactive oxygen species (ROS) levels were examined using 2'7'-dichlorodihydroflur-rescein diacetate (DCFH-DA) and the apoptosis of L cells were tested by AnnexinV-FITC/PI.Results AGEs increased thc cxpression of RAGE in a dose dependent manner.Treatment with AGEs induced a significant increase in the expression of p22phox,p47phoxand the activity of ROS,caused up-regulation of Bax and down-regulation of Bcl-2,which enhanced the apoptosis of GLUTag cells.Apocynin,the inhibitor of NADPH oxidase,prevented those responses and the effects caused by AGEs were abolished by inhibition of RAGE activity with siRNA.Conclusion AGEs positively regulate the exprcssion of NADPH oxidase-derived ROS and its down-steam signaling pathway p53/Bax by targeting RAGE,leading to the apoptosis of GLUTag cells.

In the present study, a new compound named 17-(6-cinnamamido-hexylamino-)-17-demethoxygeldanamycin (CDG) was obtained by introducing the cinnamic acid (CA) group into the 17-site of geldanamycin (GDM). The anti-cancer effects of CDG in vitro and in vivo were evaluated. MTT assay was used to examine the inhibitory effect of CDG on the proliferation of MCF-7, HepG2, H460 and SW1990 cells. Immunofluorescent staining flow cytometry combined with Annexin V-FITC/PI staining were used to detect apoptotic cells. Transwell assay was used to analyze the effect of CDG on cell invasion and migration ability. Western blotting was used to detect the expression levels of RAF-1, EGFR, AKT, CDK4 and HER-2 of MCF-7, HepG2 and H460 cells. The toxicities of CDG and GDM were evaluated in mice. Using the subcutaneously transplanted MCF-7 xenograft in nude mice, inhibitory effect was evaluated in vivo. The results showed that CDG inhibited the proliferation of cancer cells (IC50: 13.6-67.4 microg.mL-1). After exposure to CDG for 48 h, most cells presented typical morphologic changes of apoptosis such as chromatin condensation or shrunken nucleus. The rates of apoptosis of MCF-7, HepG2, H460 and SW1990 cells incubated with 10 microg.mL-1 CDG were 23.16%, 27.55%, 22.21%, 20.47%, respectively. A dose-dependent reduction of migration of four cell lines was found after exposure to CDG. The decreased levels of RAF-1, EGFR, AKT, CDK4 and HER-2 showed that CDG possessed HSP90 inhibitory effect. The result of animal toxicity test on the mice suggested that CDG had lower toxicity than GDM. Meanwhile, CDG inhibited the growth of MCF-7 xenografts of athymic mice.
Animals ; Antineoplastic Agents ; chemical synthesis ; chemistry ; pharmacology ; Apoptosis ; drug effects ; Benzoquinones ; chemical synthesis ; chemistry ; pharmacology ; Cell Line, Tumor ; Cell Movement ; drug effects ; Cell Proliferation ; drug effects ; Cyclin-Dependent Kinase 4 ; metabolism ; Female ; HSP90 Heat-Shock Proteins ; antagonists & inhibitors ; Humans ; Lactams, Macrocyclic ; chemical synthesis ; chemistry ; pharmacology ; Male ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Neoplasm Invasiveness ; Neoplasm Transplantation ; Proto-Oncogene Proteins A-raf ; metabolism ; Proto-Oncogene Proteins c-akt ; metabolism ; Random Allocation ; Receptor, Epidermal Growth Factor ; metabolism ; Receptor, ErbB-2 ; metabolism ; Tumor Burden ; drug effects ; Xenograft Model Antitumor Assays

AIMTo study the mechanisms of sodium ferulate (SF) on inhibition of collagen synthesis in hepatic stellate cells.

METHODSCollagen synthesis was analyzed by measuring 3H-proline incorporation. ELISA method was used to study the effect of SF on transforming growth factor beta1 (TGFbeta1) level in cultured HSC-T6 cell. The effect of SFon the TGFbeta1 activity in the supernatant of culture was analyzed by mink lung epithelial cell (Mv1Lu) proliferation inhibition by MTT assay.

RESULTSSF inhibited collagen synthesis in hepatic stellate cells stimulated with TGFbeta1. SF was shown to decrease TGFbeta1 level in the supernatant of HSC-T6 increased by oxidative stress. TGFbeta1 activity was intervened by SF.

CONCLUSIONSF could decrease collagen synthesis, with mechanism may be associated with that SF intervened TGFbeta1 activity, and reduced the level of TGFbeta1 increased by oxidative stress.

Animals ; Cell Proliferation ; drug effects ; Cells, Cultured ; Collagen ; biosynthesis ; Coumaric Acids ; pharmacology ; Epithelial Cells ; cytology ; Hepatocytes ; cytology ; metabolism ; Lung ; cytology ; Mink ; Oxidative Stress ; Rats ; Transforming Growth Factor beta ; metabolism ; Transforming Growth Factor beta1

Objective To investigate the therapeutic effect and safety of the clip with the flossmethod during endoscopic submucosal dissection for early gastric angle cancer. Methods A total of 27 gastric angle lesions diagnosed as early gastric cancer were treated by ESD. They were randomized to two groups, routine ESD group and clip with the flossgroup. The procedure time, complication events, en-block resection rate and complete resection rate were compared between the two groups. Also,the learning time was divided to two stages and the learning curve was studied according to the resected specimen areas per minute. Results The en block rate was 85. 7%(12/14) in the routine ESD group and 100. 0%(13/13) in the clip with the floss group. The procedure time in the clip with the flossgroup was significantly less than that in the routine ESD group (the median time 30 min VS 40 min, P =0. 011) . Perforation and the post operative bleeding did not occur in either group. The ESD learning curving during the first learning period and the mean resected specimen (area/min) in theclip with the floss group were larger than routine ESD group(30±6 mm2/min VS 20±5 mm2/min,P=0. 01). However, no difference presented during the second learning period between the two groups. Conclusion Clip with the flossmethod during endoscopic submucosal dissection for early gastric angle cancer as a novel procedure is safe, efficacious and worthy to recommend to beginning learners.

Objective To evaluate the feasibility of composite material of bone morphogenetic protein (BMP) and PLGA/TCP in repair of peri-porous-titanium bone defects in adult rabbits.Meth- otis The composite of PLGA/TCP scaffold with bovine BMP was made and implanted in distal bone de- fects peri-porous-titanium in femur of adult rabbits.The effect of BMP with PLGA/TCP on peri-prosthesis was evaluated by means of scanning electron microscope (SEM),energy dispersive X-ray analysis (EDX) and biomechanics.Results BMP with PLGA/TCP showed obvious peak of Ca and P of EDX in the pores of titanium in experiment group six weeks after operation and higher than that in control group (P＜0.05).Push-out test demonstrated that the bonding strength between the composite of HA coating/ porous titanium and bone was increased significantly with time (P＜0.05).In experiment group,at 6 and 12 weeks,peri-porous-titanium had higher shearing force compared with control group (P＜0.05). Conclusion BMP loaded with PLGA/TCP is a promising bone graft for bone defects in revision arthro- plasty,as indicates that bone induction of BMP plays an important role in biological stabilizaiton.

Objective To evaluate the feasibility and diagnostic accuracy of CE-MRA with low dose contrast agent by comparison with DSA in diabetic patients with peripheral arterial diseases. Methods ( 1 )Study in vitro: test tubes containing Gd-DTPA of different concentrations were scanned, and the relationship between signal intensities and concentrations of GD-DTPA was analyzed. DSA and CE-MRA with selected concentrations of Gd-DTPA were performed on stenotic vascular models to estimate the proper low dose of GD-DTPA for clinical applications. (2) Clinical applications: 78 diabetic patients with peripheral arterial diseases were scanned from the abdomen and pelvis station to the calf-foot station in a 3 T MR system with standard bolus chase 3D CE-MRA sequence after injection of 13 ml GD-DTPA . The image quality,diagnostic rate of stenosis of arteries in calf and degree of venous contamination were evaluated with Fisher's exact test. DSA images of 220 vascular segments in 22 patients ( 10 segments per patient) were acquired as the gold standard and compared with CE-MRA by using Kappa test. Results The MR signal intensities were proportional to the concentrations of contrast agent in present study, and all stenotic segments of vascular model were displayed by CE-MRA with GD-DTPA at lower concentration of 1.5 mmol/L. As for MRA images of 78 diabetic patients with low dose Gd-DTPA, about 97.4% (76/78) showed diagnostic image quality for pelvic and thigh stations. But the MRA images of lower extremities were interfered by the venous contamination significantly (P ＜ 0.01 ). Compared with DSA for 22 patients, the diagnostic sensitivity, specificity and agreement coefficient (Kappa value) of MRA were 96. 0% ( 168/175), 73.3%(33/45), and 0.72 (P＜0.01), respectively. Conclusion Using 3.0 T MR scanner, high quality CE-MRA of lower limb arteries can be obtained for clinical applications with contrast agent dose as low as 13 ml,which has comparable diagnostic sensitivity and specificity with DSA. But the limitation of venous contamination in MRA image should be resolved in further studies.