Objective To investigate the cause of tumor cell migration by comparing the effect of substrate stiffness on hepatic and hepatoma carcinoma cell migration so as to understand the invasive characteristics of tumor cells. Methods Immunofluorescence staining, morphological analysis and transwell were employed to observe the morphological characteristics of HCCLM3 and L02 cells on different substrates and test their migration characteristics with the quantitative analysis. Results (1) The migration rate and net translocation of HCCLM3 and L02 cells on 4 kPa substrate was higher than those both on 0.5 kPa(most soft one) and on glass (the hardest one) substrates, and L02 cells also displayed higher migration efficiency than HCCLM3 cells on such substrates. (2) The mean squared displacement of HCCLM3 and L02 cells on different substrates showed consistent tendency, and the directional persistence of L02 cells on the softer substrate was significantly higher than that of HCCLM3 cells. (3) In 0.5 and 1 mg/mL three dimensional collagen environment, the number of invasive cells of HCCLM3 was remarkably more than that of L02 cells. After adding MMPs inhibitor GM6001 (40 μg/mL), the number of invasive cells was notably increased in HCCLM3 cells, but notably decreased in L02 cells. Conclusions (1) In two dimensional comparatively soft environment, L02 cells displayed an efficient migration due to its higher directional persistence. (2) In three-dimensional collagen environment, the invasion efficiency of HCCLM3 cells was significantly higher due to the various modes of migration adaptation to the microenvironment.

Background Proliferation of the human Tenon capsule fibroblasts(HTFs) is a main cause of failure of filtering surgery.To search the drug of inhibiting the growth of the HTFs is essential for the improvement of successful rate of filtering surgery.Objective The aim of this study was to investigate the inhibitory effect of apigenin on HTFs and its mechanism.Methods Human Tenon capsular tissue was obtained during the strabismus correction surgery.HTFs was primarily cultured using explant method and identified using vimentin by immunochemistry.The 3-5 generation of cells were incubated to 96-well plate.Apigenin of 0,20,40,80,160 μmol/L was added into the medium,respectively,for 24,48,72 hours,and the proliferation of HTFs was detected by sulfonyl chloride (SRB) at the wavelength of 560 nm (A560).Bromodeoxyuridine (BrdU) of 10 μg/L was added to culture the cells for 48 hours to calculate the labeling rate of BrdU.The morphology of the cells was observed using Hoechst 33258 staining,and apoptosis and cells cycle were evaluated by flow cytometry.Results Cultured cells grew well with the positive response for vimentin,showing the green fluorescence in cytoplasm.SRB assay showed that the A560 value was gradually declined with the increase of the dosage of apigenin and prolong of time (Fgroup =480.306,P =0.000 ; Ftime =555.144,P =0.000).The labeling rate after 0,40,80 μmol/L apigenin acted for 48 hours was (87.860 ±0.632)％,(61.520±4.306)％ and (23.480±4.472)％,showing a significant difference among the three groups (F =299.347,P =0.000).The labeling rate of HTFs for BrdU was significantly decreased in the 40 and 80 μmol/L apigenin groups compared with the 0 μmol/L apigenin group (P＜0.05).Hoechse 33258 staining found that the number of the HTFs was gradually decreased and the cell number of karyopyknosis and nuclear deformation was increased with the increase of apigenin dosage.Percentage of cells in G0/G1 phase were raised and that in S and G2/M phase were declined in the higher dosage apigenin group,with a significant difference among the different groups (FG0/G1 =58.621,P=0.000;Fs =32.357,P=0.001 ;FG2/M =83.998,P=0.000).In the 72nd hour after acted by 0,40,80,160 μmol/L apigenin,the apoptosis rate of HTFs was (4.77±0.21) ％,(13.24±1.35)％,(18.33±1.86) ％,(31.58 ± 2.77) ％,respectively,with a statistically significant difference among the four groups (F =204.791,P＜0.05).Conclusions Apigenin restrains the growth of HTFs by evoking G0/G1 cell cycle arrest and inducing apoptosis in a dosage-and time-dependent manner.

Objective To evaluate the value of multimodal MRI in differential diagnosis of lung squamous cell carcinoma and adenocarcinoma.Methods Routine sequence,diffusion weighted imaging(DWI)and dynamic enhancement images about 1 6 squamous cell carcinoma and 21 adenocarcinoma patients were analyzed retrospectively.Taken a record about the size,edge,internal signal, enhancement types and apparent diffusion coefficient(ADC)values when b=600 s/mm2,and the difference in the degree of pathological differentiation was studied.Results The average diameter of squamous cell carcinoma was (4.17±2.0)cm,while adenocarcinoma was (3.81±1.8)cm,lobulated and spiculation were the most common signs in both of them.Squamous cell carcinoma showed low T1signal in 12 cases(75%),low T2signal in 7 cases(43.7%),adenocarcinoma showed high T1signal in 10 cases(47.6%),high T2 signal in 14 cases(66.7%).Squamous cell carcinoma had lower ADC value than adenocarcinoma(1.27×10-3mm2/s vs 1.38×10-3mm2/s), and well differentiated tumors had higher ADC values than poor ones,it was statistically significant.The most common time-signal intensity curves were type A in squamous cell carcinoma(62.5%)and type B in adenocarcinoma(50%).Conclusion MRI findings of squamous cell carcinoma and adenocarcinoma are associated with the biological characteristics,squamous cell carcinoma has shorter T2signal and adenocarcinoma has shorter T1signal.Squamous cell carcinoma has lower ADC value than adenocarcinoma and is dominated by outflow curve (type A),these features are helpful in subtype and differential diagnosis.

OBJECTIVETo provide the basis for clinical acquired immunodeficiency syndrome (AIDS) surveillance and to avoid cross infection in hospital, we study the infection status of AIDS in Shandong province.

METHODSThe fourth-generated Akzo's ELISA kit and the fourth generated Immunoluminometric detection reagent were used for HIV antibody screening for 399 303 cases of both inpatients and outpatients from Jan. 2003 to Dec. 2011. Beijing WanTai ELISA kit and Se-marked rapid detection reagent were used for re-detection, and the positive samples were sent to the local CDC for confirming test by Western Blot.

RESULTSThe HIV-1 antibody detection results of 129 (0. 3230 per thousand) patients were confirmed to be positive, including 54 (0. 1352 per thousand) cases of outpatients and 75 (0. 1878 per thousand) cases of inpatients. HIV infection rates in outpatients from 2003 to 2011 were 0.050 per thousand, 0.030 per thousand, 0.111 per thousand, 0.120 per thousand, 0.124 per thousand, 0.113 per thousand, 0.148 per thousand, 0.201 per thousand, 0.2152 per thousand; and that in inpatients were 0. 150 per thousand, 0.089 per thousand, 0.138 per thousand, 0. 144 per thousand, 0. 104 per thousand, 0. 132 per thousand, 0. 197 per thousand, 0. 329 per thousand, 0. 313 per thousand respectively. Among these inpatients, there were 61 cases of medical patients and 14 cases of surgical patients, and most were youths and farmers.

CONCLUSIONSHIV infection rate was increasing year by year. Most inpatients whose HIV-1 antibody was positive were in the phase of AIDS. Therefore, it's very necessary to execute routine testing for inpatients and outpatients who need special examination for early diagnosis of HIV infection

Acquired Immunodeficiency Syndrome ; epidemiology ; immunology ; virology ; Adolescent ; Adult ; Aged ; China ; epidemiology ; Enzyme-Linked Immunosorbent Assay ; Female ; HIV Antibodies ; analysis ; HIV Infections ; epidemiology ; immunology ; virology ; Humans ; Male ; Middle Aged ; Young Adult

OBJECTIVETo study the mechanism of Ca(2+) on the apoptosis induced by hyperthermia in neonate rat hippocampal neurons to provide the applicative evidence of dantrolene for preventing brain injuries.

METHODSDantrolene, Ca(2+) specific blocking agent, was used in the hyperthermia-induced apoptosis of primary hippocampal neurons in vitro to observe its effect on the apoptosis, fluorescent intensity, and dynamic change of Ca(2+) by flowcytometry and laser confocal microscopy.

RESULTSThe rate of apoptosis was decreased significantly after hyperthermia treatment by dantrolene sodium. The intracellular Ca(2+) fluorescent intensity in 42 degrees C treatment group (107.35 +/- 6.0) was significantly lower than that in control group (159.12 +/- 33.8). The concentration of Ca(2+) began to decrease 20 approximately 25 s after adding dantrolene sodium, and reached the lowest level about 50 s later, and then kept lower than the basal level.

CONCLUSIONDantrolene sodium has an important protective effect on hippocampal neurons apoptosis induced by hyperthermia and may have some applicative value of preventing heat-induced brain injury.

Animals ; Animals, Newborn ; Apoptosis ; drug effects ; Calcium ; metabolism ; Calcium Channel Blockers ; pharmacology ; Cells, Cultured ; Dantrolene ; pharmacology ; Female ; Hippocampus ; cytology ; drug effects ; Male ; Neurons ; drug effects ; metabolism ; Rats ; Rats, Sprague-Dawley ; Temperature

OBJECTIVETo evaluate the toxicity of chicken calamus keratin (CCK) conduit as a tissue-engineered scaffold material.

METHODSThe chemical composition of the leaching solution of CCK was determined by means of ultraviolet spectrometry, and the toxic effects of the solution was evaluated by skin sensitization test in rats, intracutaneous stimulation test in rabbits, acute systemic toxicity test in mice, and cytotoxicity test in L929 cells.

RESULTSThe leaching solution of CCK consisted mainly of middle-molecular-weight peptides with a small quantity of macromolecular proteins. Skin sensitization test in rats showed that application of the CCK leaching solution caused no obvious skin reddening, regional edema, or skin necrosis. Intracutaneous injection of the leaching solution in rabbits did not induce obvious skin stimulation manifested by intradermal erythema or edema. In acute systemic toxic test, administration of the leaching solution in mice caused no death, organ dysfunction, cyanosis, tremor, severe peritoneal irritation, ptosis, or dyspnoea. In vitro cytotoxicity test indicated that the cell toxicity of the CCK leaching solution was approximately at 0 level.

CONCLUSIONCCK contained in the treated chicken calamus easily undergoes hydrolysis to release mainly some peptides which do not induce obvious toxic effects, suggesting the safe potential applications of CCK conduit as a tissue-engineering biomaterial.

Animals ; Cell Line ; Cell Proliferation ; drug effects ; Chickens ; Feathers ; chemistry ; Female ; Keratins ; chemistry ; toxicity ; Male ; Mice ; Rabbits ; Rats ; Skin Irritancy Tests ; Solutions ; Tissue Engineering ; Tissue Scaffolds ; chemistry ; Toxicity Tests ; methods

Objective Toexploretheimagingcharacteristicsofileocecaltuberculosisbycontrast-enhancedCT.Methods TheCT findingsof16ileocecaltuberculosispatientswereretrospectivelyanalyzed.AllcasesunderwentplainandenhancedCTscan.Results 14casesshowedinhomogeneousthickeningoftheintestinalwallandnarrowingofthelumen,and2casesshowedextensiveanduniform thickeningoftheintestinalwallwithnoobviousnarrowingofthelumen.Theenhancedimagesshowedthat13caseswereobviously heterogeneousenhancement,2caseswerering-likeenhancement,and1casewashomogeneousenhancement.Blurredserosasurface wasseeninallpatients,withsurroundingexudationin14casesandperitoneuminvolvementin12cases.Inallcases,enlargedlymph nodeswerefoundneartheabdominalaortaandonmesentery,withhomogeneousenhancementin10cases,annularenhancementin5casesand calcificationin1case.Conclusion Ileocecaltuberculosisismainlycharacterizedbyinhomogeneousthickeningandheterogeneousenhancementofthe intestinalwall,aswellasnarrowingofthelumen.Thereisoftenobviousexudationaroundtheintestines.Ileocecaltuberculosisoften combineswithenlargedlymphnodesandintrapulmonarytuberculosis.TheenhancedCTscanisofgreatvaluefortheevaluationofileocecal tuberculosis.

Anal Canal/surgery* ; Humans ; Rectal Fistula/surgery* ; Treatment Outcome

OBJECTIVETo develop a composite material containing human hair keratin (HHK), collagen sponge (inner layer) and poly 2-hydroxyethyl methacrylate (PHEMA) film that allows sustained release of polydatin and test its effect as a biological dressing in promoting burn wound healing in SD rats.

METHODSThree HHK materials with fast, moderate, and low degradation rates were mixed at the ratio of 4:3:3 to prepare a reticular structure, which was processed into a composite material with bovine tendon-derived collagen sponge, and further complexed with HEMA film containing PD prepared by polymerization. Degree II burn wound was induced in SD rats by scalding and within postburn day 2-5, the wounds were cleansed and covered with the composite material or with glutaraldehyde-treated porcine skin (positive control). At week 1, 2, 4, 6 and 8 following wound dressing, 6 full-thickness skin samples were harvested from the wounds for histological observation and immunohistochemical detection of collagen and elastic fibers, and the wound healing time and healing rate were recorded.

RESULTSThe prepared collagen sponge film was transparent and porous (50-300 microm in diameter) and allowed sustained PD release into normal saline within 48 h. Compared with the porcine skin, the composite material reduced exudation and maintained ideal moisture of the wound, and significantly shortened the wound healing time (P=0.000). On day 7, 14, and 21 following dressing, the composite material and porcine skin significantly increased the wound healing rate as compared with the negative control group (P=0.000), and on day 14, the composite achieved significantly greater healing rate than the porcine skin (P<0.05).

CONCLUSIONHHK-collagen sponge-PHEMA/PD composite as a dressing material promotes burn wound healing in rats by allowing in vivo construction of tissue engineered epidermis. PHEMA is feasible for sustained drug delivery in this composite.

Animals ; Biological Dressings ; Burns ; drug therapy ; Cattle ; Collagen ; therapeutic use ; Drug Delivery Systems ; Drugs, Chinese Herbal ; pharmacology ; Glucosides ; pharmacology ; Humans ; Keratins ; therapeutic use ; Polyhydroxyethyl Methacrylate ; therapeutic use ; Rats ; Rats, Sprague-Dawley ; Stilbenes ; pharmacology ; Swine ; Tissue Engineering ; Wound Healing

OBJECTIVETo improve the histocompatibility of chicken calamus keratin (CCK) graft by collagen-gel coating or using of cyclosporine A (CsA).

METHODSThirty SD rats were equally randomized into 5 groups, and in 4 of them, CCK implantation into the bilateral erector spinae was performed on different treatment protocols. In group A, the rats received daily intraperitoneal injection of CsA (5 mg/kg) for two consecutive weeks after CCK implantation; in group B, CCK was soaked in CsA (2.5 mg/ml) solution at 4 degrees Celsius; for 48 h before grafting; in group C, CCK coated with collagen gel was grafted; and in group D, only CCK was implanted. Rats in the fifth group received only cutaneous incision as well as muscular dissection to serve as the blank control. CCK degradation and its effect on the surrounding tissues were observed at 2, 4 and 8 weeks after grafting. Immunohistochemistry was performed to identify T lymphocyte infiltration in the host tissues.

RESULTSAll the rats survived the operation. Numerous macrophages, especially multinucleated giant cells occurred on the peripheral of the CCK grafts, and small degraded CCK pieces were observed in their cytoplasm. Only a few inflammatory cells were seen in the host tissues. At 2, 4 and 8 weeks after CCK implantation, only a few CD3-positive cells were found in all the groups, and in group A and B, the density of T lymphocytes was significantly lower than that in group D, and there was no significant difference between group A and the blank control group.

CONCLUSIONSCsA significantly improves the histocompatibility of CCK material, and short-term systemic CsA administration achieves the best results. Macrophages, especially multinucleated giant cells participate in CCK degradation in vivo.

Animals ; CD3 Complex ; analysis ; Chickens ; Coated Materials, Biocompatible ; administration & dosage ; chemistry ; Collagen ; chemistry ; Cyclosporine ; administration & dosage ; chemistry ; Feathers ; chemistry ; Female ; Gels ; Histocompatibility ; drug effects ; Immunohistochemistry ; Immunosuppressive Agents ; administration & dosage ; chemistry ; Implants, Experimental ; Injections, Intraperitoneal ; Keratins ; chemistry ; Male ; Muscle, Skeletal ; chemistry ; drug effects ; surgery ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Spine ; T-Lymphocytes ; chemistry ; cytology ; Tissue Engineering ; methods