Objective To investigate the effects of emodin on the migration and invasion ability and expressions of E-cadherin and Slug of human hepatoma cell lines HepG2; To discuss the possible mechanisms of anti-hepatocellular carcinoma.Methods HepG2 cells were cultured in vitro. The experimental group was treated with emodin at concentrations of 10μmol/L and 20μmol/L as. The negative control group was treated with the same volume of RPMI-1640 medium, while the positive control group was treated with 10μmol/L floxuridine. The cell matrix adhesion assay, wound healing and transwell chamber in vitro invasion assay were used to observe the effects of emodin on HepG2 cell adhesion rate and migration and invasion ability. Western blot analysis was used to observe the changes of expressions of E-cadherin and Slug.Results Compared with the negative control group, emodin inhibited significantly HepG2 cell adhesion rate and migration and invasion ability were in a dose-dependent manner (P<0.05,P<0.01); Western blot analysis showed that the protein expression of E-cadherin increased significantly, and the level of Slug decreased significantly in a dose-dependent manner (P<0.05,P<0.01).Conclusion Emodin can significantly inhibit migration and invasion of HepG2 cells, which mechanism may up-regulate expressions of E-cadherin and down-regulate Slug.

[Objective] To recommend an ideal method for extracting Ⅰ-collagen from cortical bone.[Method]The cortical bone of pig was splitted into small pieces after the soft tissues were cleaned up.The bone pieces were gradually dehydrated in alcohol,defatted in aether,decalcificated in hydrochloric acid,redefatted in chloroform:methanol(1:1,v/v)and became soft.The soft pieces were disintegrated into demineralized bone matrix(DBM)powder in a high speed mill.The osseins were extracted respectively by enhanced pepsin digestion method or alkali-solution method after the DBM powder was treated with desolving,centrifuging,dialyzing and lyophilization.The product got by enhanced pepsin digestion method was further confirmed.The extraction rate and appearance,viscosity and solidification of both products were analyzed and compared.[Result]The collagen produced by extraction of enhanced pepsin digestion method was confirmed to be I-collagen by analyzing amino-acid composition,protein electrophoresis,relative moleculas weight and max wavelength about light absorption.The extraction rate of Ⅰ-collagen by enhanced pepsin digestion method was(94.0?14.96)% as comparision with by alkali-solution method(57.8?4.96)% and the viscosity test of acetic and solution at the concentration 0.03%(w/v)was 3.71 and 2.81 respectively.At the condition of 37?,pH 7.35~7.45,the collagen solution extracted by enhanced pepsin digestion method solidified and changed into a glue 10 minutes later,while the product solution by extraction with alkali-solution was still like a sticky liquid.[Conclusion]Ossein extracted from cortical bone with enhanced pepsin digestion method is a realⅠ-collagen.Compared with alkeli-solution extraction method,the enhanced pepsin digestion extraction method has advantages of higher extraction rate,better purity,better viscosity and fine solidification of the product,and is a choice to prepare the Ⅰ-collagen from cortical bone.

[Objective]To investigate the expressions of CD-147 in osteosarcoma samples and to research clinically pathological significance. [Methods]The expressions of CD-147 was detected by immunohistochemistry SABC method in 55 cases of osteosarcoma samples.[Results]The moderate/strong expression of CD-147 was 54.5% compared with giant cell tumor of bone.And the high expressions of CD-147 was in close correlation with diagnosis and prognosis in osteosarcoma.[Conclusion]CD-147 were overexpressed in osteosarcoma,which can be relevant to the malignant progression of osteosarcoma.

OBJECTIVE:To optimize the extracting process of the total flavonoids from Herba Epimedii.METHODS: The extract technology of the total flavonoids from Herba Epimedii was optimized by orthogonal experiment with the yield of total flavonoids from Herba Epimedii as index and with the extraction temperature,extraction time,the ratio of solid to liquid and the concentration of ethanol as factors.RESULTS:The optimum extracting conditions for the total flavonoids from Herba Epimedii were as follows:the reflux extraction was conducted twice(1 hour/time) at 80 ℃ with 60% ethanol as solvent,and the ratio of solid to solution was 1∶10.CONCLUSION:The optimum extracting technology is simple and reasonable,and it serves as a good reference for the extracting process of the total flavonoids from Herba Epimedii.

BACKGROUND:Human umbilical cord mesenchymal stem cells are similar to bone marrow mesenchymal stem cells, which can directional y differentiate into neuron-like cells, secrete various cytokines, and provide the base for nerve regeneration. ＜br> OBJECTIVE:To study the role of chitosan composite nerve conduit carrying umbilical cord mesenchymal stem cells in nerve end-to-side anastomosis. ＜br> METHODS:Thirty while rabbits were randomized into three groups. The central branch of the right posterior peroneal nerve were cut and proximal y ligated, and then sutured evaginably to the muscle. In the control group, the distal end of the common peroneal nerve were anastomosed into the tibial nerve at 30°-45°;in the stenting group, the chitosan conduit was bridged at the same interval and angle into the end-to-side anastomosis site between the tibial nerve and peroneal nerve;in the cel-stenting group, the chitosan conduit carrying human umbilical cord mesenchymal stem cells was bridged at the same interval and angle into the end-to-side anastomosis site between the tibial nerve and peroneal nerve. After 12 weeks, gross observation, neurophysiological examination and anti-S-100 immunohistochemistry detection were performed. ＜br> RESULTS AND CONCLUSION:After 12 weeks of operation, in the cel-stenting group, the conduit degraded completely, the nerve diameter was similar to that of the normal peroneal nerve, and the motor nerve conduction velocity was higher than that in the control and stenting groups (P＜0.01). Anti-S-100 immunohistochemistry results showed that a great amount of brownish red Schwann cells arranged around the regenerated nerve fibers in the cel-stenting group, while there was few and sparse brownish red substance, and the Schwann cells grew worse in the stenting and control groups. These findings suggest that umbilical cord mesenchymal stem cells have an obvious role in promoting nerve regeneration, induce bud growth, accelerate the growth rate of regenerated fibers, and improve growth and maturity of Schwann cells.

AIM: To investigate the antitumor effects of tumor necrosis factor- ? (TNF - ?) and interferon -?(IFN -?) on hepatocellular carcinoma (HCC) . METHODS: Cytotoxicity of the combination of TNF-? and IFN-? on HCC in vitro was measured by using a crystal violet (CV) staining method. Antitumor effects of the combination of TNF- ? and IFN - ? on HCC in vivo were observed by intra - hepatic injection of TNF-? and IFN-? to the tumor in a human HCC nude mice hepatic model. RESULTS: The growth of HCC cells was inhibited by TNF -? alone, which was dose - dependent. The cytotoxicity of TNF -? on HCC was enhanced by incubation with IFN -?. TNF at 107 U/L, or IFN -? at 106 U/L alone killed only 27.1 % or 7.9 of HCC cells, respectively, when combined with IFN -?, it killed 83.7% of HCC cells. A synergistic antitumor effect on HCC in vivo was observed in combination group, as tumor growth inhibition rate was 35.9% compared with 17.2% in TNF-? group and 5.6% in IFN -? group. The survival period of mice bearing tumor was significantly prolonged and serum AFP was significantly decreased in combination group (P

BACKGROUND: Umbilical cord blood mesenchymal stem cells (UCB-MSCs) can differentiate into various types of cells under certain condisions, and easily proliferate in vitro. However, UCB-MSCs have long establishment time and low frequency.OBJECTIVE: To in vitro isolate and culture UCB-MSCs, and induce its differentiation into osteoblasts.DESIGN, TIM E AND SETTING: The in vitro cytological study was performed at the Laboratory of the Medical College of Qingdao University from June 2008 to January 2009.MATERIALS: UCB was obtained from term normal delivery women at the Department of Gynaecology and Obstetrics, Qingdao Municipal Hospital.METHODS: Human UCB-MSCs were isolated and cultured in vitro by Percoll density gradient. When reached 90% confluency,UCB-MSCs were digested by trypsin for subculture. At the third passage, UCB-MSCs at 1×106 were incubated. When reached 50% 60% cenfluency, UCB-MSCs were treated with DMEM supplemented with 0.1 μmol/L dexamathasone, 10 mmol/Lβ-sodium glycerophosphate and 50 μmol/L vitamin C. UCB-MSCs in the control group were incubated in low glucose DMEM.MAIN OUTCOME MEASURES: Growth and proliferation of MSCs were observed under the inverted microscope. Cell surface marker expression and cell growth curve were measured by flow cytometry. Cell ultrastructure was observed under the transmission electron microscope. Differentiation of UCB-MSCs into osteoblasts was determined by Won Kossa staining and alkaline phosphatase staining.RESULTS: Primary cultured UCB-MSCs had similar morphology to bone marrow mesenchymal stem cells. After passage, cell morphology was even, presenting spindle shape. UCB-MSCs at passage 3 highly expressed CD29, CD44, CD13, but did not express CD34. Growth latency was 2-3 days. Cells entered logarithm proliferation phase at days 3-4, and platform phase 1 month later. Nuclei presented round or irregular, with clear nuclear membrane, 1-2 nucleoli, rough chromatin, abundant organelles and microvilli. UCB-MSCs at passage 3 were gradually confluent following 3 days of osteogenic induction, with the presence of pavement-stone shape. 14 days later, calcified nodules by Von Kossa staining, and cells were positive for alkaline phosphatase staining. In the control group, no calcified nodules were found, and cells were negative for alkaline phosphatase staining.CONCLUSION: UCB-MSCs can be successfully isolated by Percoil density gradient, and induced to differentiate into osteoblasts in vitro.

BACKGROUND: Previous studies have successfully prepared the natural and biologically degraded acellular nerve graft and have proved the effect of promoting neural regeneration.OBJECTIVE: To construct tissue engineered artificial nerve with acellular nerve graft and bone marrow mesenchymal stem cells, and to observe the effect of promoting motor functional recovery and repairing rat sciatic nerve defects. DESIGN, TIME AND SETTING: Randomized control animal experiment was performed in the Medical TIssue Engineering Laboratory of the First Affiliated Hospital of Liaoning Medical University between June 2008 and February 2009. MATERIALS: Wistar adult healthy male rats weighing 180-200 g were used to prepare acellular nerve graft, while Wistar adult healthy male rats weighing 100-120 g were used to prepare bone marrow mesenchymal stem cells. Tissue engineered artificial nerve was produced with acellular nerve graft co-cultured with bone marrow mesenchymal stem cells. METHODS: Sixty Wistar adult healthy male rats weighing 180-200 g were induced sciatic nerve defect models, 15 mm long. SD rats were divided into three groups at random with 20 animals in each group. ①Experiment group: Rat sciatic nerve defects were bridged with tissue engineered artificial nerve. ②Blank control group: Rat sciatic nerve defects were bridged with tissue engineered nerve scaffold. ③Autologous nerve control group: Rat sciatic nerve defects were bridged with autologous nerve graft. MAIN OUTCOME MEASURES: At 12 weeks postoperation, the recovery of motor function was evaluated with gross observation, electrophysiology, histological observation and triceps surae wet weight.RESULTS: ①At 12 weeks postoperation, the toes at the operation side could separate and supported to the ground in the experiment group; there was no significant difference in the regenerated nerve conduction velocity between experimental group and autologous nerve graft group. ②At 12 weeks postoperation, histochemical stain results showed AchE-positive motor end-plate arranged regulady in the middle and superior part of gestrocnemius muscle to form end-plate zone in the experiment group. By use of silver staining, the regenerated nerve tract and the emergent branch were shown to be connected with motor end-plate.③There was no significant difference in the tibialis anterior muscle wet weight between experimental group and autologous nerve graft group. CONCLUSION: Bridging acellular nerve graft and bone marrow mesenchymal stem cells into rat sciatic nerve defects can promote motor functional recovery.

BACKGROUND:Biological dressing A (porcine Xenoderm) and Physiotule Ag (Coloplast) show good effects on the absorption of exudates, adhesion and anti-bacteria in treatment of degree Ⅱ facial burns. OBJECTIVE:To observe the effectiveness of biological dressing Aversus Physiotule Ag after early debridement of degree Ⅱ facial burns. METHODS:A total of 15 patients with superficial degree Ⅱ facial burn and 10 patients with deep degree Ⅱ facial burns were included. Symmetric facial area of the same patient with the same depth of burn was divided into two parts of same size for treatments. One side was randomly selected as the experimental side, and treated with biological dressing A. The other was selected as the control side and treated with Physiotule Ag. We compared wound healing time, infection, times of changing dressings, skin after healing, drug change comfort and dressing comfort between the two sides. RESULTS AND CONCLUSION:In patients with superficial grade Ⅱ facial burn, times of changing dressings and drug change comfort were better in the experimental side than in the control side (P < 0.05), but the dressing comfort was better in the control side than the experimental side (P< 0.05). No significant difference in wound healing time, infection and skin after healing was detected between the two sides. In patients with deep degree Ⅱ facial burns, wound healing time, times of changing dressings, skin after healing and drug change comfort were better in the experimental side than in the control side (P < 0.05), but the dressing comfort was better in the control side than the experimental side (P< 0.05). No significant difference in infection was detectable between the two sides. Above findings suggested that the therapeutic effects of biological dressing A and Physiotule Ag were similar in treatment of degree Ⅱ facial burns. Biological dressing A in repair of deep degree Ⅱ facial burns promotes the wound healing andelevates the quality of healing.

BACKGROUND:The interaction of neural network and motor function in post-stroke brain tissue remains unclear.
OBJECTIVE:To observe neural network impairment fol owing subacute stroke by using diffusion tensor imaging, and to investigate the relationship with neurological defects and motor dysfunction.
METHODS:A total of 19 patients after subactue stroke and 20 healthy adults were examined with diffusion tensor imaging. The fol owing parameters were compared:fractional anisotropy, apparent diffusion coefficient, asymmetry indices of fractional anisotropy and apparent diffusion coefficient. The neurological defect and motor function were evaluated with the corresponding scales. The 10-meter walking speed was measured. The correlation of diffusion tensor imaging parameters with the scale scores and 10-meter walking speed was analyzed.
RESULTS AND CONCLUSION:The stroke group exhibited reduced fractional anisotropy value asymmetry and fractional anisotropy value in bilateral posterior limbs of the internal capsule. Apparent diffusion coefficient value asymmetry and apparent diffusion coefficient value in the posterior limb of the internal capsule were lower than control unaffected side (P<0.05). Apparent diffusion coefficient value and apparent diffusion coefficient value asymmetry in posterior limb of the internal capsule showed a strong negative correlation with Fugl-Meyer assessment scores of the lower extremities (P<0.05). Diffusion tensor imaging parameters is closely linked with motor dysfunction of the lower extremities in subacute stroke patients. Local stroke lesion-caused neurological defect is the leading cause of motor dysfunction of the lower extremities.