Synthesis of Heterocyclic Ring-Fused Bisnoralcohol Derivatives as Novel Small-Molecule Antiosteoporosis Agents.

A series of heterocyclic ring-fused derivatives of bisnoralcohol (BA) were synthesized and evaluated for their inhibitory effects on RANKL-induced osteoclastogenesis. Most of these derivatives possessed potent antiosteoporosis activities in a dose-dependent manner. Among these compounds, 31 (SH442, IC50 = 0.052 µM) exhibited the highest potency, displaying 100% inhibition at 1.0 µM and 82.8% inhibition at an even lower concentration of 0.1 µM, which was much more potent than the lead compound BA (IC50 = 2.325 µM). Cytotoxicity tests suggested that the inhibitory effect of these compounds on RANKL-induced osteoclast differentiation did not result from their cytotoxicity. Mechanistic studies revealed that SH442 inhibited the expression of osteoclastogenesis-related marker genes and proteins, including TRAP, TRAF6, c-Fos, CTSK, and MMP9. Especially, SH442 could significantly attenuate bone loss of ovariectomy mouse in vivo. Therefore, these BA derivatives could be used as promising leads for the development of a new type of antiosteoporosis agent.

[Anatomic study and application of TRAM flap with partial preservation of abdominal rectus muscle in the breast reconstruction].

OBJECTIVE: To investigate the feasibility and effectiveness of transverse rectus abdominis musculocutaneous (TRAM) flap with partial preservation of abdominal rectus muscle based on the anatomic study in cadavers. METHODS: 5 adult female cadavers which provided by department of anatomy of Fujian Medical University were dissected after injection with medical red latex from the starting point of the inferior epigastric artery and superior epigastric artery. The TRAM flap with partial preservation of lateral abdominal rectus muscle were dissected for breast reconstruction. The location, route, branches and anastomosis of inferior and superior epigastric arteries were observed. Based on the anatomic study, breast reconstruction were performed in 8 cases with muscle-sparing TRAM flaps. RESULTS: The inferior epigastric artery arises from external iliac artery (9/10, 90%) or femoral artery (1/10, 10%) at the joint point between the internal third and lateral two third. There are extensive anastomoses between superior and inferior epigastric arteries above the umbilicus, mostly between the 2cm below the first tendinous intersection and umbilical level. From Sept. 2009 to Sept. 2010, 8 cases received breast reconstruction with muscle-sparing TRAM flap. The patients were followed up for 3 months to one year. Fibrosis happened in subcutaneous fat at flap IV zone in 2 cases, borderline necrosis and subcutaneous fat liquefaction occurred in some areas of flap IV zone in 2 cases, which healed after debridement. The other 4 cases healed with no complication. Except for unsatisfied shape in one case, good result achieved in 7 cases. There was no abdominal weakness, hemia or other complication. CONCLUSIONS: It is an effective and safe method in breast reconstruction with muscle-sparing TRAM flap. It is practical with comparatively short operation time and less morbidity in donor site.

[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.

[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.

[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 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.

[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.