BACKGROUND:CD133 has been used as a marker to separate and study different tumor stem cells by using its protein specific features. OBJECTIVE:To investigate the expression of stem cell surface marker CD133 in hepatocel ular carcinoma and the in vitro proliferation of its positive subsets. METHODS:The human hepatoma MHCC97-H cell lines were cultured and sorted, CD133+and CD133-MHCC97-H cells were obtained, and the expression of CD133 and cell migration and invasion were detected. After 7 days of culture, cell proliferation ability and Notch expression were detected. After 14 days of culture, cell cloning efficiency was detected. CD133+and CD133-MHCC97-H cells were implanted into nude mice subcutaneously, and 4 weeks later, tumor formation in mice was detected. RESULTS AND CONCLUSION:(1) CD133 expression and cell clone formation:The expression of CD133 and cell cloning efficiency of CD133+MHCC97-H cells were significantly higher than those of CD133-MHCC97-H cells (P<0.05). (2) Cell proliferation:After 3-7 days of culture, the absorbance value of CD133+MHCC97-H cells was significantly higher than that of CD133-MHCC97-H cells (P<0.05). (3) Cell migration and invasion:The number of CD133+MHCC97-H cells passing through the cell membrane was significantly more than that of CD133-MHCC97-H cells (P<0.05). (4) The Notch protein expression of CD133+MHCC97-H cells was significantly higher than that of CD133-MHCC97-H cells (P<0.05). (6) Tumor formation test:The tumor size of CD133 -MHCC97-H cells was larger than that of CD133-MHCC97-H cells (P<0.05). To conclude, CD133+hepatocel ular carcinoma cell subsets with strong proliferation ability have some characteristics of cancer stem cells, and strong invasion, metastasis and tumorigenic abilities.

Objective: To study the toxic effects of 5-amionlevulinic acid-based photodynamic therapy (ALA-PDT) on human peripheral blood mononuclear cells (PBMCs), cord blood mononuclear cells (CBMCs) and peripheral blood stem cells (PBSCs), and furthermore, to understand the possible causes of this response. Methods: We used MTT assay to detect the survival rate of PBMCs, CBMCs and PBSCs after treated by ALA-PDT under the optimum experiment conditions with U937 as control; Annexin V-FITC/PI was used to detect the pattern of cell death induced by ALA-PDT. By using flow cytometry, we detected intracellular PpIX fluorescence intensity. Results: After ALA-PDT treatment the survival rate of PBMCS had no significant change; however in PBSCS and CBMCs, the survival rate reduced to 70%, and the survival rate of leukemia cell U937 was the lowest, about 30%. After incubation with ALA, except for PBMCs, intracellular PpIX fluorescence intensity of the other two kinds of normal haemocytes and U937 increased obviously. These results combined with the flow cytometry suggested that the main pattern of cell death here was apoptosis. Conclusion: Under the optimum experiment conditions, ALA-PDT has a slight effect on normal haemocytes but excellent depletions of leukemia cells. Therefore, it can effectively purify autologous bone marrow or stem cell grafts.

Objective: To study the toxic effects of 5-amionlevulinic acid-based photodynamic therapy (ALA-PDT) on human peripheral blood mononuclear cells (PBMCs), cord blood mononuclear cells (CBMCs) and peripheral blood stem cells (PBSCs), and furthermore, to understand the possible causes of this response. Methods: We used MTT assay to detect the survival rate of PBMCs, CBMCs and PBSCs after treated by ALA-PDT under the optimum experiment conditions with U937 as control; Annexin V-FITC/PI was used to detect the pattern of cell death induced by ALA-PDT. By using flow cytometry, we detected intracellular PpIX fluorescence intensity. Results: After ALA-PDT treatment the survival rate of PBMCS had no significant change; however in PBSCS and CBMCs, the survival rate reduced to 70%, and the survival rate of leukemia cell U937 was the lowest, about 30%. After incubation with ALA, except for PBMCs, intracellular PpIX fluorescence intensity of the other two kinds of normal haemocytes and U937 increased obviously. These results combined with the flow cytometry suggested that the main pattern of cell death here was apoptosis. Conclusion: Under the optimum experiment conditions, ALA-PDT has a slight effect on normal haemocytes but excellent depletions of leukemia cells. Therefore, it can effectively purify autologous bone marrow or stem cell grafts.

Objective To optimize experimental parameters for the photosensitization of 5-aminolevulinic acid (ALA) in promyelocytic leukemia cell HL60 and compare them with normal human peripheral blood mononuclear cell (PBMC). Methods ALA incubation time, wavelength applied to irradiate, concentration of ALA incubated, irradiation fluence may modulate the effect of 5-aminolevulinic acid based Photodynamic Therapy (ALA-PDT).The high-pressure mercury lamps of 400W served as light source, the interference filter of 410nm, 432nm, 545nm, 577nm were used to select the specific wavelength. Fluorescence microscope was used to detect the fluorescence intensity and location of protoporphyrin IX (PpIX) endogenously produced by ALA. MTT assay was used to measure the survival of cell. Flow cytometry with ANNEXIN V FITC kit (contains annexin V FITC, binding buffer and PI) was used to detect the mode of cell death. Results ① 1mmol/L ALA incubated 1×105/mL HL60 cell line for 4 hours, the maximum fluorescence of ALA induced PpIX was detected in cytomembrane. ② Irradiated with 410nm for 14.4J/cm2 can result in the minimum survivability of HL60 cell. ③ The main mode of HL60 cell death caused by ALA-PDT is necrosis. Conclusion ALA for 1mmol/L, 4 hours for dark incubation time, 410nm for irradiation wavelength, 14.4J/cm2 for irradiation fluence were the optimal parameters to selectively eliminate promyelocytic leukemia cell HL60 by ALA based PDT. The photosensitization of ALA based PDT caused the necrosis of HL60 cell, so it could be used for inactivation of certain leukemia cells.

Objective To understand the acute effect of the air pollution on daily mortality of cardiovascular diseases in the old people aged over 65 years in Taiyuan, Shanxi, China. Methods Case-crossover design was used to assess the association between air pollution and daily cardiovascular disease mortality. Pollutants considered were nitrogen dioxide (NO2), carbon monoxide (CO), inhalable particles (PM10) and sulfur dioxide (SO2). Results CO exposure was significantly associated with the mortality of five kinds of cardiovascular diseases. No association was found for arrhythmia. As CO concentration increased by 100 ?g/m3, the corresponding OR of cardiovascular disease mortality were as follows, total cardiovascular disease mortality were1.000-1.013, cardiac mortality were1.000-1.024, ischemic heart disease were 1.005-1.032, cardiac failure were 1.006-1.112, myocardial infarction were 1.009-1.050 respectively. The higher OR value was found in unidirectional control design than that in the bidirectional design. Considering the lag effect of air pollution, the 24 hr and 48 hr average of air pollution levels were analyzed in the present study respectively. Conclusion The current air pollution level (PM10, CO) in Taiyuan City has a acute effect on the cardiovascular disease mortality in the local old people aged over 65 years.

Objective The growth inhibitory effects of GlcNH_2?HCl,GlcNH_2 and NAG on human hepatoma SMMC-7721 cells in vitro was investigated.The antitumor activity of GlcNH_2?HCl against Sarcoma_(180) in KM mice was also investigated.Methods The cell viability in vitro was examined with MTT assay.DNA isolation and cell-cycle analyses were also performed.GlcNH_2?HCl was ig administered to Sarcoma_(180) KM mice.The inhibition rates,spleen and thymus index were calculated.Results GlcNH_2?HCl and GlcNH_2 resulted in a concentration-dependent reduction in hepatoma cell growth.The inhibition rates against SMMC-7721 cell of GlcNH_2?HCl and GlcNH_2 at concentration of 500?g?mL~(-1) were(50.24)% and 52.19%.As to the concentration of 1000?g?mL~(1), the rates were(82.21%) and 83.20%.This effect was accompanied by a marked increase in the proportion of S phase cells.Compared with the control,there was no significant difference among various concentrations of NAG,GlcNH_(2)?HCl exhibited inhibitiory effect against Sarcoma_(180) in mice at the dosage of 125～500 mg?kg~(-1),and the inhibition rate was about 27.84%～34.02%.The optimal inhibitory effect was 250 mg?kg~(-1).GlcNH_2?HCl could enhance the weights of thymus and spleen.In addition,it could promote lymphocyte transformation.Conclusion It is therefore postulated that the antitumor effect of GlcNH_2?HCl is probably host-mediated and cytocidal.

Objective:To further study concentration-response relationship between daily mortality and particulate air pollution,to provide scientific basis for making policy.Methods:We applied spline model to daily time-series data for Taiyuan city,China from 2004-2005,using concentration of particulate matter less than 10 ?m in aerodynamic diameter(PM10)as the exposure measure,and controlling confounding factors,such as season,meteorological factors,day-of-the-week,other air pollutions,etc.Results:The spline model showed a non-linear relationship for the relative risks of death for all causes(total deaths)and for specific causes in relation to PM10 concentration,and obtained threshold levels.For total mortality,malignant tumor mortality and cardiovascular-cerebrovascular-respiratory mortality,the threshold levels were 85.00 ?g/m3,89.59 ?g/m3,122.54 ?g/m3,respectively,using the value of differential deduction.Conclusion:Our nonlinear models are appropriate for assessing the effect of particulate air pollution on daily mortality at current ambient levels in Taiyuan,China,in contrast to those of Europe and America.

Objective To observe the effect of stellate ganglion block (SGB) in the treatment of peri-men-opanse syndrome of clinical efficacy. Methods 30 patients diagnosed as perimenepausal syndorme by the gynecolo-gy clinic in our hospital from February 2007 to December 2008 were selected. All patients experienced vaginal cytolo-gy and examination of blood estradiol (E2),follicle-stimulating generation Su (FSH),luteinizing hormone (LH),in line with perimenopausal syndrome and no other chronic diseases, and in the last 3 months the patients had not taken hormone treatment drugs. Anterior SGB once a day,around the turn was adopted,taking 10 times as a course of treat-ment. All patients were treated for two courses. The blood FSH, LH, E2 changes were recorded. Results Blood E<,2> in-creased from (31.29±19.36) pmol/L to (159.47±88.21) pmol/L(t=-24.976, P<0.01). FSH decreased from (54.67±19.24) U/L to (38.15±13.50) U/L (t=13.872, P<0.01), and LH dropped from (36.1± 15.6)U/L to (26.7±8.7)U/L (t=9.188,P<0.01). Conclusion SGB has the disorder and autonomic Endo-crine function to achieve a new balance because it can adjust perimenopausal autonomic nervous imbalance, so it is the effective treatment for elimination of peri-menopause syndrome.

Objective The visual inspection method were not appropriate to perform a hemolysis evaluation for colored injection like doxorubicin hydrochloride,this article adopted three methods to evaluate the hemolysis test of doxorubicin hydrochloride in vitro and provide reference for clinical drug safety.Methods Using rabbit erythrocytes as experimental object,the durg concentration 4.0 and 2.0 mg/mL was chosen which range of clinical concentration and preclinical safety evaluation concentration,to evaluate the hemolysis test of doxorubicin hydrochloride injection with blood analyzer test,direct colorimetric assay,and indirect colorimetric assay.Results The evaluation results of three different methods were very consistent.The tube's hemolysis rate of 4.0 mg/mL dose was far greater than 5％,which means serious hemolysis;Only 0.1 mL tube of 2.0 mg/mL dose (according to the drug concentration equal to 0.5 mL tube of 0.4 mg/mL drug concentration) without hemolysis occurring,the other tubes' hemolysis rates were far greater than 5％,which means serious hemolysis.Conclusion The hemolysis phenomenon may occur when 2.0 mg/mL dose of doxorubicin hydrochloride solution for iv injection is used in clinic and dilution (final concentration not more than 0.4 mg/mL) is recommended.

Objective To study the toxic effects of 5-amionlevulinic acid-based photodynamic therapy (ALA-PDT) on human peripheral blood mononuclear cells (PBMCs), cord blood mononuclear cells (CBMCs) and peripheral blood stem cells (PBSCs), and furthermore, to understand the possible causes of this response. Methods We used MTT assay to detect the survival rate of PBMCs, CBMCs and PBSCs after treated by ALA-PDT under the optimum experiment conditions with U937 as control;Annexin V-FITC/PI was used to detect the pattern of cell death induced by ALA-PDT. By using flow cytometry, we detected intracellular PpIX fluorescence intensity. Results After ALA-PDT treatment the survival rate of PBMCs had no significant change;however in PBSCs and CBMCs, the survival rate reduced to 70%, and the survival rate of leukemia cell U937 was the lowest, about 30%. After incubation with ALA,except for PBMCs, intraceUuiar PplX fluorescence intensity of the other two kinds of normal haemocytes and U937 increased obviously. These results combined with the flow cytometry suggested that the main pattern of cell death here was apoptosts. Conclusion Under the optimum experiment conditions, ALA-PDT has a slight effect on normal haemocytes but excellent depletions of leukemia cells. Therefore, it can effectively purify autologons bone marrow or stem cell grafts.