[Photochemotherapy with psoralen and ultraviolet A induced apoptosis of NB4 cells and its effects on caspase-8 and caspase-8 protein expressions].

OBJECTIVE: To study the regulatory effects of psoralen (PSO) plus ultraviolet A (UVA), which is PUVA, on cell apoptosis of human leukemia cell line NB4 and signal pathway of cell apoptosis. METHODS: Human leukemia cell line NB4 was cultured in vitro. The NB4 cells were treated with PSO extracted from Chinese medicine psoralea fruits at different concentrations (0, 5, 10, 20 and 40 microL) plus UVA of wave length 360 nm at different irradiation time points (0 and 5 min). The apoptosis ratio was detected by flow cytometry (FCM). The ultrastructure changes were observed using transmission electron microscope (TEM). The expressions of Caspase-8 and Caspase-8 protein were detected by immunocytochemical method (ICC). RESULTS: After treatment of PSO at different concentrations with a 0 and 5-min exposure of UVA, the apoptosis rate of NB4 cells increased dose-and time-dependently, and was up to peak after treatment of PSO at 40 microg/mL with 5-min exposure of UVA. An interaction was shown between the two factors (P <0. 01). There were obvious morphological apoptosis of NB4 cells under TEM after treated with PUVA. The expressions of Caspase-3 and Caspase-8 protein were up-regulated by PSO, UVA, and PUVA, but the effects of PUVA on Caspase-3 protein were stronger than PSO and UVA at 12 h time-dependently (P <0.01).An interaction was shown between the concentration of PSO and time of UVA (P <0.01). CONCLUSIONS: The optimal combination of PUVA was PSO in 40 microg/mL and 5-min exposure of UVA. PUVA could induce the apoptosis of NB4 cells and in vitro activate Caspase-3 and Caspase-8 genes.

[Effect of combined application of psoralen and ultraviolet A for inducing NB4 cell apoptosis and its impact on Fas/FasL gene expressions].

OBJECTIVE: To explore the effect of combined application of psoralen (PSO, an extract from psoralea) and ultraviolet A (UVA) for inducing the apoptosis of human leukemic cell line NB4 and its impact on the Fas/FasL gene expressions. METHODS: According to factorial design, changes of apoptosis rate and ultrastructure of NB4 cells, as well as the gene and protein expressions of Fas/FasL were observed after cells were treated with PSO in different concentrations and irradiated by UVA of 360 nm wavelength for different times, using flow cytometry, transmission electron microscopy and quantitative polymerase chain reaction (PCR), and the outcomes were treated with variable analysis. RESULTS: (1) After treatment of PSO in concentration of 10, 20, 40, 80 microg/mL combined with a 5-min exposure of UVA, the NB4 cells apoptosis rate induced were 26.57% +/- 0.42%, 30.67% +/- 0.11%, 34.90% +/- 0.30% and 24.63% +/- 0.38% respectively. The effects were dose- and time-dependent, and an interaction was shown between the two actors. (2) After being treated by PSO plus UVA, obvious ultrastructure changes with apoptosis characteristics were shown in NB4 cell under electron microscope. (3) PSO plus UVA showed up-regulatory effect on gene and protein expressions of Fas, and down-regulatory effect on gene and protein expressions of FasL in a dose- and time-dependent manner, with the interaction between the two actors in altering Fas gene expression, also in altering FasL gene and protein expressions. CONCLUSION: Combined application of PSO and UVA can induce the apoptosis of NB4 cells, and the Fas/FasL system is one of the pathways for apoptosis inducing.

[Compound huangdai tablet as induction therapy for 193 patients with acute promyelocytic leukemia].

OBJECTIVE: To report the results of curative and adverse effects of compound huangdai tablet (CHDT) as induction therapy for 193 patients with acute promyelocytic leukemia (APL). METHODS: CHDT was administered 1.25 g orally three times a day after meal for three days, then the dosage was gradually increased to 7.5 g/d. RESULTS: One hundred and ninety-three patients achieved complete remission (CR), 78.8% of whom in 30 to 60 days with an average time of 44.3 d. No serious infection, bleeding or DIC occurred during the treatment course. The major adverse effects were gastrointestinal symptoms. There was no change in lanine transaminase, urea, creatinine or electrocardiographic QTc interval in 110 APL patients observed before and after the treatment. CONCLUSION: CHDT therapy is a modality of higher CR rate, good safety and tolerance without bone marrow suppression for APL patients.

[Effects of PUVA on expression of Fas and FasL in inducing apoptosis of NB4 and K562 cells].

OBJECTIVE: To explore the effects of psoralen (PSO) and long wave ultraviolet A (PUVA) on expression of Fas and FasL in apoptosis of NB4 and K562 leukemia cells. METHODS: The NB4 and K562 cells were taken as the study objects and their apoptosis ratios, ultrastructure changes and the expression of Fas and FasL were detected in order to observe the effects of PSO and UVA of wave length 360 nm on human leukemia cells. The factorial design and analysis of variance were used to analyze the interaction among the factors. RESULTS: All of PSO, UVA and PUVA could induce the apoptosis of NB4 and K562 cells, and the effects of PUVA were stronger than the other two. After treated with PUVA, the NB4 and K562 cells all showed obvious ultrastruture changes about apoptsis under the electron microscope. All of PSO, UVA and PUVA could increase the expression of Fas gene and protein, and decrease the expression of FasL gene and protein. Moreover, the effects of PUVA were stronger than the other two. CONCLUSION: PUVA can induce the apoptosis of NB4 and K562 cells and the effects are the strongest, one of the pathway of PUVA to induce apoptosis is to upregulate the expression of Fas gene and downregulate the expression of FasL gene.

[Effect of psoralen plus longwave UVA inducing HL-60 cells apoptosis].

The aim of this study was to investigate the effects of the traditional Chinese medicine psoralen (PSO) plus long wave ultraviolet A (PUVA) on apoptosis in HL-60 leukemia cells and its mechanism. The effect of PUVA on HL-60 cell growth was assayed by MTT method and the changes of ultrastructure of cells were observed by electron microscopy. The apoptosis ratios, changes of mitochondrial membrane potential, expression of Fas and FasL protein and Fas and FasL mRNA were detected by FCM and fluorescent quantitation RT-PCR respectively. The expression of Caspase 8 and Caspase 3 protein were detected by immunocytochemistry (ICC). The results showed that the growth of HL-60 cells were inhibited by PUVA in time-and concentration-dependent manner through inducing cell apoptosis. When the irradiation time of long wave ultraviolet A lasted 15 minutes and the concentration of PSO was 80 microg/ml, the inhibition of HL-60 cell proliferation and apoptosis ratios reached the peak. There were obvious apoptotic ultrastructure changes and decrease of mitochondrial membrane potential in HL-60 cells after treatment with PUVA. The expression of Fas mRAN increased and expression of FasL mRNA decreased after treating with PUVA for 4 hours, and the same results of Fas, FasL expression on protein level were obtained also after treating with PUVA for 24 hours. The expression of caspase 8 and caspase 3 protein enhanced and reached the peak after treating with PUVA for 8 hours. It is concluded that the PUVA can inhibit the growth of HL-60 cells and induce apoptosis of these cells. The possible mechanism is supposed to be up-regulating Fas, down-regulating FasL levels and then activating the levels of caspase 8 and caspase 3. The decreasing of mitochondrial membrane potential may be involved in this process probably.

[Effects of psoralen with long wave ultraviolet light on apoptosis and mitochondrial membrane potential in K562 cells].

OBJECTIVE: To study the effects of psoralen (PSO) with long wave ultraviolet light (PUVA) on apoptosis and mitochondrial membrane potential in K562 cells. METHODS: K562 cells were incubated with PSO in different concentrations (10, 20, 40 and 80 microg/ml for 24 hours) and irradiated without or with UVA (5 min). The changes in ultrastructure of the cells were observed under a transmission electron microscope. The apoptosis rates and the changes of mitochondrial membrane potentials were detected by flow cytometry. The factorial design and analysis of variance were used to analyze the interaction among the factors. RESULTS: There were obvious ultrastructure changes related to apoptosis in K562 cells after being treated with PUVA (80 microg/ml). PSO, UVA and PUVA all increased the apoptosis rates and decreased the mitochondrial membrane potentials, and the effects of PUVA were stronger than those of PSO and UVA (P<0.01). CONCLUSION: PUVA can induce the apoptosis of K562 cells and one of the pathways to the induction of apoptosis is to down-regulate the mitochondrial membrane potentials.

[Effects of psoralen plus long-wave ultraviolet-A on apoptosis of HL-60 leukemia cells].

OBJECTIVE: To explore the effects of psoralen (PSO) plus long-wave ultraviolet-A (PUVA) on apoptosis and expression of Fas ligand (FasL) in HL-60 leukemia cells. METHODS: The HL-60 cells were taken as the study objects and their apoptosis rates, ultrastructure changes and the expression of FasL were detected in order to observe the effects of PSO and ultraviolet-A (UVA) of wave length 360 nm. The factorial design and analysis of variance were used to analyze the interaction among the factors. RESULTS: PSO, UVA and PUVA all induced the apoptosis and the effects of PUVA were stronger than those of the other two. After HL-60 cells had been treated with PUVA, they all showed obvious ultrastructure changes due to apoptosis observed under the electron microscope. PSO, UVA and PUVA all decreased the expressions of FasL gene and protein. The effects of PUVA were stronger than those of the other two. CONCLUSIONS: PUVA can induce the apoptosis of HL-60 cells and the effects are stronger than those of PSO or UVA alone. The expression of FasL gene in HL-60 cells is down-regulated during the apoptosis induced by PUVA.

[Progress in studies of mechanism of anti-fibrosis traditional Chinese drugs and effective components].

The progress in the research of pharmacological mechanism of anti-fibrosis traditional Chinese drugs and the effective components is summarized. It's demonstrated by pharmacological experiments that anti-fibrosis traditional Chinese drugs can inhibit the cell proliferation, regulate the cytokines and ECM, and intervente in the signal transduction.

[Effects of PUVA on mitochondrial membrane potential in HL-60 and K562 cells].

OBJECTIVE: To study the effects of psoralen (PSO) and long wave ultraviolet light (PUVA) on mitochondrial membrane potential in HL-60 and K562 cells. METHODS: Cells were incubated with PSO in different concentrations irradiated with or without UVA. The changes of ultrastructure of cells were observed under the electron microscope. The apoptosis ratios and the changes of mitochondrial membrane potential were detected through the flow cytometry. The factorial design and analysis of variance were used to analyze the interaction among the factors. RESULTS: There were obvious ultrastructure changes about apoptosis in HL-60 and K562 cells after treated with PUVA. PSO, UVA and PUVA all increased the apoptosis ratios and decreased the mitochondrial membrane potential, and the effects of PUVA were stronger than the other two (p<0.01). CONCLUSION: PUVA can induce the apoptosis of HL-60 and K562 cells and one of the pathway to induce apoptosis is to downregulate the mitochondrial membrane potential.

[Effects of inactivated rabbit serum containing compound realgar and natural indigo tablet on cell line NB4].

OBJECTIVE: To explore the effects of inactivated rabbit serum containing compound realgar and natural indigo tablet (CRNIT) on cell line NB(4). METHODS: The experimental rabbits were taken as the provider of the animal serum, and the serum was inactivated before the experiment. The serum was divided into two groups based on whether the rabbits were given CRNIT. The concentration of arsenic in the rabbit's serum was detected by AFS-230a double path atom fluorescence photometer. The inhibition rates and apoptosis rates were regarded as the observational indexes. RESULTS: The concentration of arsenic in the inactivated rabbit serum containing and not containing the drug were (0.010 0+/-0.001 0) mg/L and (0.110 0+/-0.006 4) mg/L respectively, and the difference had statistical significance (P<0.01). The two groups of serum all had inhibitory effect on the growth of NB(4) cells depending on the drug concentration and effect time. And there were significant differences among the groups. The two groups of serum all induced the apoptosis of NB(4) with positive relations with the concentration and effect time. And there were significant differences among the groups. CONCLUSIONS: The rabbit serum containing CRNIT can obviously restrain the growth of NB(4) cells and the inhibitory effect depends on the concentration and effect time. And the rabbit serum containing CRNIT can also induce the apoptosis of NB(4) cell line and the apoptosis rates depend on the concentration and effect time.

[Growth-inhibiting effect of psoralen plus ultraviolet-A light therapy on K562 cells].

OBJECTIVE: To observe the effects of psoralen plus ultraviolet-A light (PUVA) on K562 cells and the relative mechanism. METHODS: The effects of psoralen, ultraviolet-A light and PUVA on K562 cells were assayed by monotetrazolium test (MTT). DNA content was analyzed by flow cytometry (FCM). The apoptotic rates of K562 cells treated with 40 and 80 microg/ml psoralen for 24 and 48 hours were assayed by Annexin-V-FITC/PI reagent kit on FCM respectively. The ultrastructures of apoptotic cells were observed by a transmission electron microscope (TEM). RESULTS: Either single psoralen therapy or single ultraviolet-A irradiation had inhibiting effect on K562 cells. The inhibiting effect of PUVA on K562 cells was stronger than that of the single psoralen therapy or single ultraviolet-A light irradiation (P<0.05). Apoptotic peak (AP) was detected by FCM. TEM test showed that K562 cells treated with PUVA were smaller, having condensed cell nucleus, assembled chromatin, disintegrated nucleus body and the majority of the cells appeared to be apoptotic conformation. CONCLUSION: Psoralen has inhibiting effect on K562 cells, and the effect of PUVA is more significant. It is suggested that 10 min irradiation and 40 microg/ml terminal concentration of psoralen be probably the best choice for PUVA. The inhibiting effect of PUVA is due to apoptosis.

[Study on Realgar inducing apoptosis in T lymphocytic cell line CEM].

OBJECTIVE: To study the Realgar induced T lymphocytic leukemia cell line CEM apoptosis in vitro. METHODS: CEM cells were incubated with Realgar. Cell proliferation inhibition was determined by MTT. Cell cycle, apoptosis, Apo2.7 and Fas were measured by cytometer. RESULTS: Realgar inhibited the proliferation of CEM cell line. The cells treated with Realgar showed a Sub-G(0)/G(1) apoptotic peak in DNA distribution histogram, increment of Apo2.7 protein expression, and arrested cells in G(2)/M phase, but ineffectiveness on Fas expression. CONCLUSION: The Realgar can induce CEM cell apoptosis in vitro.

[Department of a finite element model of the Head Model of HYBRID III Dummy with the Human Mandible].

OBJECTIVE: To analyze the biomechanics of impact injury in the condition of the simulate impact on mandible. METHODS: A finite element model of human mandible was developed from the CT scan images by the technologies of three-dimensional reconstruction, image processing and meshing. The mandible model was connected to a modified head model of HYBRID III dummy with joint according to the anatomic structure and mechanical characteristics of the temporomandibular joint. RESULTS: A finite element model of human head with true anatomic structure mandible has been developed. This model has been validated with the cadaver test results. The higher stress was showed in the condyle rejoins and coracoid in the model when mandible was in impact simulation. CONCLUSIONS: This model can be used to research the mechanism of craniofacial blunt-impact injury and assess the injury severity. The model of HYBRID III dummy with the human mandible was helpful for the boundary design of mandibular model in the impact simulations.