BackgroundBiological behavior of choroidal melanoma is closely related with angiogenesis.The microcirculation pattern in which tumor cells may be involved is different from neovascularization.ObjectiveThis study was to observe the microcirculation pattern of the choroidal melanoma tissue and analyze its relationship with the clinical pathology factor and the degree of cellular proliferation.Methods Forty-eight specimen of choroidal melanoma tissue were collected at Beijing Tongren Eye Centre from November 1997 through March 2006.Periodate acid Schiff staining was used to determine the microcirculation pattern of choroidal melanoma.The morphology of the tumor cells and the distribution of microcirculation pattern in the tumor were observed under a fluorescence microscopy with 544 nm wavelength,and the Ki-67 expression in the tumor was detected by immunochemistry.The relationships between the choroidal melanoma clinical pathology factor with Ki-67 expression and microcirculation pattern were evaluated by multiple stepwise regression model.ResultsA total of 9 kinds of microcirculation patterns were found by periodate acid Schiff staining.The occurring rate of loop- or network-like vascular pattern showed a significant elevation in the tumor tissue with epithelial cells in comparison with the tumor with spindle-like cells (66.7％ vs.30.6％ ) ( P=0.027 ).The expression rates of Ki-67 were 18.961 ± 10.995 and 19.481 ± 12.167 in the tumor tissue with loop- or network-like microvascular pattern,and those in the tumor tissue without loop- or network-like microvascular pattern were 10.261 ±5.669 and 12.021 ± 6.802,presenting significant differences between them ( P =0.000,P =0.010).Loop-like microvascular pattern was determined as the risk factor of the proliferation and metastasis of choroidal melanoma by multiple stepwise regression analysis (P=0.002).ConclusionsAmong the nine microcirculation patterns in choroidal melanoma,networks pattern is the most fashion,and Ki-67 expression is more strong in the tumor with epitheliod cells,suggesting that this microcirculation pattern is associated to the malignant degree and extent of proliferation in choroidal melanoma.

The shape and structure of granules are controlled by the granulation process, which is one of the main factors to determine the nature of the solid dosage forms. In this article, three kinds of granules of a traditional Chinese medicine for improving appetite and promoting digestion, namely, Jianwei Granules, were prepared using granulation technologies as pendular granulation, high speed stirring granulation, and fluidized bed granulation and the powder properties of them were investigated. Meanwhile, synchrotron radiation X-ray computed micro tomography (SR-µCT) was applied to quantitatively determine the irregular internal structures of the granules. The three-dimensional (3D) structure models were obtained by 3D reconstruction, which were more accurately to characterize the three-dimensional structures of the particles through the quantitative data. The models were also used to quantitatively compare the structural differences of granules prepared by different granulation processes with the same formula, so as to characterize how the production process plays a role in the pharmaceutical behaviors of the granules. To focus on the irregularity of the particle structure, the box counting method was used to calculate the fractal dimensions of the granules. The results showed that the fractal dimension is more sensitive to reflect the minor differences in the structure features than the conventional parameters, and capable to specifically distinct granules in structure. It is proved that the fractal dimension could quantitatively characterize the structural information of irregular granules. It is the first time suggested by our research that the fractal dimension difference (Df,c) between two fractal dimension parameters, namely, the volume matrix fractal dimension and the surface matrix fractal dimension, is a new index to characterize granules with irregular structures and evaluate the effects of production processes on the structures of granules as a new indicator for the granulating process control and optimization.
Drugs, Chinese Herbal ; analysis ; Fractals ; Medicine, Chinese Traditional ; Powders ; Quantitative Structure-Activity Relationship ; Synchrotrons ; Technology, Pharmaceutical ; Tomography, X-Ray Computed

Objective To observe the relationship between apoptosis of K562 cell induced by iron-deprivation and activation of Caspase-3.Methods K562 cells were treated with desferrioxamine(DFO) in different dosages were collected at different time points.K562 cells were labelled with Annexin V/PI,and then the rate of apoptosis was measured by flow cytometry;The activation of Caspase-3 were detected by colorimetric method with pAN labelled substrate;The active protein of Caspase-3 were analyzed by Western blot.Results When K562 cells were treated with different concentrations of DFO,the apoptosis rate and the activity of Caspase-3 increases gradually.When K562 cells were incubated with DFO(50 ?mol/L and 100 ?mol/L) 24 h later,the enzymatic activity of Caspase-3 increases dramatically more than that of control group,and the difference was significantly(P0.05).All those effect above can be counteracted by equal mole concentration of FeCl_3.Conclusion Iron-deprivation maybe induce the apoptosis of K562 cell by chelating intracellular iron and activing Caspase-3.

Objective To investigate the effects of smad7 on transdifferentiation and collagenⅠsynthesis in advanced glyeosylation end-products(AGE)-stimulated NRK52E cells.Methods NRK52E cells were transferred by pTet-on plasmid system and the cell lines of doxycycline(Dox)-regulated Smad7 expression were selected for the study.Transnuclear location of p-Smad2/3 was examined with immunocytochemistry.The mRNA and protein expressions of Smad7,?-SMA,E-cadherin,collagenⅠwere detected with RT-PCR and Western blot. Results AGE-induced expressions of Smad7 mRNA and protein were further increased in NRK52E cells by the addition of Dox in a dose-dependent manner.Overexpression of Smad7 caused a marked inhibition of p-Smad2/3 transnuclear location at 30 min(68.3% vs 31.2%,P

Background Choroidal melanoma (CM)is the most common primary intraocular tumor,and brachytherapy is one of the most common therapeutic modality in the treatment of the tumor.However,this irradiation approach has not been evaluated in China. Objective The present study was to analyze the effectiveness and safety of domestic 125I plaque irradiation in the treatment of CM. Methods Forty New Zealand albino rabbits were randomized into 5 groups with 8 rabbits 8 eyes (right eyes) in each group.CM models were established in 16 of 40 New Zealand albino rabbits by implanting the rat B16F10 melanoma cell fragments into the suprachoroidal space of right eyes.After 3 weeks,domestic 125I plaque was fixed at the location of CM in the irradiation group 1,and 8 rabbits with CM served as model control group.The clinical effectiveness of 125I plaque for CM was evaluated based on the fundus examination with indirect ophthalmoscopy,B scan ultrasonography,fundus photographs and color Dopplerimaging.Regarding the safety study,domestic 125I plaque was fixed on the normal right cycs of normal rabbits,while the plaques without 125I seeds were used as the sham group.No intervene was performed in the rabbits of blank group.The number of CD4+,CD8+ T cells in peripheral blood was detected by flow cytometry before plaques implanted and on 3,7,15 and 30 days after the plaque was removed.The animals were sacrificed and the eyes were obtained for histology examination.The use of the experimental animals complied with Statement of ARVO. Results After implantation of B16F10 melanoma cell fragments,CM grew steadily and rapidly with the similar size between irradiation group 1 and model control group ( P =0.550).One week after administration of the treatment,tumor size was(0.31±0.07 )cm in irradiation group 1 and (0.85±0.18 )em in the model control group,with the significant difference between them( P=0.001 ).Two week after application of 125I plaque,the size of tumor was smaller than that before irradiation (P=0.007 ).Histologically,the tumors were mostly limited beneath the pigment epithelial layer with less neovascularization,fibrosis in the tumor was found in some area in the irradiation group when compared with model control group.No significant differences were found in the proportions of CD4+,CD8+ T cells and CD4+/CD8+ at different time points in the irradiation groups of normal eyes and sham group (Fgroup =0.770,8.110,2.230; P=0.380,0.060,0.140; Ftime =0.770,3.220,4.230; P =0.550,0.170,0.004 ).Chronic inflammatory cells infiltration cornea,subconjunctival epithelial and selera surface,but sclera had no necrosis and organization.Conclusions These results suggest that domestic 125I plaque irradiation is effective for the treatment of CM,and has limited side effects on normal rabbits.

OBJECTIVETo investigate the effect of tissue-engineering bone on repair of segmental bone defects.

METHODSSegmental bone defect of 21mm was created at sheep left metatarsus, which was then implanted with tissue-engineering bone (the experimental group) and pure porous beta-TCP (the control group) respectively. The bone defect in the blank group was left without treatment. After the sheep were sacrificed at the 1st, 3rd, or 6th month postoperatively, the samples were taken and examined by radiological, histological and biomechanical methods as well as scanning electron microscopy. The sheep in the blank group were sacrificed at the 6th month postoperatively.

RESULTSThe osteoid tissue, woven bone and lamellar bone in the defect of the experimental group occurred earlier than in the control group. The new bone formed directly without through a cartilaginous intermediate in the experimental group, while the defect was repaired in a "creep substitution" way in the control group. At the 6th month, radiological and biomechanical tests revealed nearly complete repair of the bone defect of the experimental group, partial repair in the control group and non-healing in the blank group.

CONCLUSIONSTissue-engineering bone can repair bone defect, accelerating healing and without "creep substitution", which is a good option in repair of critical segmental bone defects. This study set up a basis for clinical applications in the future.

Animals ; Biocompatible Materials ; Bone Diseases ; surgery ; Bone Marrow Cells ; cytology ; Bone Regeneration ; Calcium Phosphates ; therapeutic use ; Cell Culture Techniques ; Cells, Cultured ; Mesenchymal Stromal Cells ; cytology ; Sheep ; Tissue Engineering ; methods ; Transplantation, Autologous

OBJECTIVETo investigate the relationship between coxsackievirus infection and type 1 diabetes mellitus (T1DM), and observe the changes of T lymphocyte subsets in the development of T1DM.

METHODSWe detected Coxsackievirus RNA by reverse transcription PCR, and measured the change in T-lymphocyte subsets by flow cytometry in 22 cases of newly diagnosed T1DM (group I), 30 patients with diabetes for some time (group II), and 30 healthy subjects (group III).

RESULTSThe positivity rate of coxsackie virus RNA in groups I, II, and III was 55.55%, 23.33%, and 6.67%, respectively, showing a significant difference among the 3 groups (P<0.01). Patients with upper respiratory tract infection had a higher positivity rate for coxsackie virus RNA than those without upper respiratory tract infection in group I (P<0.05). Compared with the control group, the percentage of CD3, CD4 and CD4/CD8 ratio decreased significantly in groups I and II (P<0.01 or P<0.05). CD3, CD4 and CD4/CD8 tended to increase in group II in comparison with group I, and there was an significant difference in CD3 and CD4 between the two groups (P<0.01 or P<0.05). Compared with the control group and CVBRNA-negative group, CVBRNA-positive group showed significantly lowered CD3, CD4, CD8 and CD4/CD8 (P<0.01 or P<0.05).

CONCLUSIONThe occurrence and development of type 1 diabetes is closely related to coxsackie virus infection, and the changes in T lymphocyte subsets serves as a probable mechanism of its pathogenicity.

Adolescent ; Adult ; CD4-CD8 Ratio ; CD4-Positive T-Lymphocytes ; immunology ; CD8-Positive T-Lymphocytes ; immunology ; Coxsackievirus Infections ; complications ; immunology ; Diabetes Mellitus, Type 1 ; complications ; virology ; Female ; Humans ; Lymphocyte Count ; Male ; T-Lymphocyte Subsets ; immunology ; Young Adult

Syncope is a common emergency of children and adolescents,which has serious influence on the quality of life.Neurally-mediated syncope,including postural tachycardia syndrome,vasovagal syncope,orthostatic hypotension and orthostatic hypertension,is the main cause of syncope in children and adolescents.The main manifestations of neurally-mediated syncope are diverse,such as dizziness,headache,chest tightness,chest pain,pale complexion,fatigue,pre-syncope and syncope.Although the clinical manifestations are similar,each subtype of syncope has its hemodynamic feature and optimal treatment option.The diagnosis rate of syncope in children has been greatly improved on account of the development of the diagnostic procedures and methods.In recent years,with the promotion of head-up tilt test and drug-provocated head-up tilt test,the hemodynamic classification of neurally-mediated syncope gets continually refined.In recent years,with the effort of clinicians,an appropriate diagnostic protocol for children with syncope has been established.The initial evaluation consists of history taking,physical examination,standing test and standard electrocardiography.After the initial evaluation,some patients could be diagnosed definitely,such as postural tachycardia syndrome,orthostatic hypotension,and situational syncope.Those with a specific entity causing syncope need selective clinical and laboratory investigations.Patients for whom the cause of syncope remained undetermined should undergo head-up tilt test.The precise pathogenesis of neurally-mediated syncope is not entirely clear.In recent years,studies have shown that neurally-mediated syncope may be related to several factors,including hypovolemia,high catecholamine status,abnormal local vascular tension,decreased skeletal muscle pump activity and abnormal neurohumoral factors.Currently based on the possible pathogenesis,the individualized treatment of neurally-mediated syncope has also been studied in-depth.Generally,the management of neurallymediated syncope includes non-pharmacological and pharmacological interventions.Patient education is the fundamental part above all.In addition to exercise training,the first-line treatments mainly include oral rehydration salts,beta adrenoreceptor blockers,and alpha adrenoreceptor agonists.By analyzing the patient's physiological indexes and biomarkers before treatment,the efficacy of medication could be well predicted.The individualized treatment will become the main direction in the future researches.

OBJECTIVETo observe the different extracts from Radix Isatidis on multiplication of mice lymphocytes.

METHODLymphocytes were separated and cultured. Immunological activities of different extracts from Radix Isatidis were studied by thermodynamics and the results were tested by the conventional pharmacological experiments.

RESULTThe results showed that the water extract and residue had significant immunological effects while organic solvent extracts had immunological activity to some extent.

CONCLUSIONThe comparison of immunological activity among the extracts from Radix Isatidis were as follows: residue after extracting > general extract > nBuOH extract > EtOAc extract > CHCl3 extract > P E extract.

Animals ; Calorimetry ; Cell Proliferation ; drug effects ; Cells, Cultured ; Drugs, Chinese Herbal ; isolation & purification ; pharmacology ; Isatis ; chemistry ; Lymphocytes ; drug effects ; Male ; Mice ; Plant Roots ; chemistry ; Plants, Medicinal ; chemistry

OBJECTIVE: To investigate the potential effect of relaxin family peptide receptor 1( RXFP1) in the process of silica-induced silicosis. METHODS: Sixty-four specific pathogen free male Wistar rats were randomly divided into control group and experimental group. By one time intratracheal infusion,rats in experimental group were treated with 0. 1 m L 500 g / L silica dust suspension while the control group was treated with 0. 1 m L sodium chloride physiological solution. Eight rats from each group were sacrificed on day 1,7,14 and 28 after exposure. Histopathologic changes of the lung tissue were performed with hematoxylin-eosin staining. The expressions of Rxfp1 mRNA and RXFP1 protein in rat lungs were detected by real-time polymerase chain reaction and immunohistochemical staining,respectively. RESULTS: After 28 days of exposure,the grey nodules were observed by naked eye in the lung of the experimental group. The fracture and silicotic nodules could be seen in alveolar interval with light microscope. Compared with the control group,the Rxfp1 mRNA relative expression level in the lungs of experimental group was increased to 145% after 1 day of exposure( P < 0. 01),followed by a decrease on day 7 and 14 and reached similar level of control group( P > 0. 05). By day 28,it dropped to45% of control group( P < 0. 01). The RXFP1 protein relative expression in experimental group was significantly up-regulated since the 7th day compared to that of the control group( P < 0. 01). And it reached to the highest level on the28 th day( P < 0. 01). CONCLUSION: The RXFP1 might play an important role in inhibiting silicosis.