Anisotropic cell growth at the leaf base promotes age-related changes in leaf shape in Arabidopsis thaliana.

Plants undergo extended morphogenesis. The shoot apical meristem (SAM) allows for reiterative development and the formation of new structures throughout the life of the plant. Intriguingly, the SAM produces morphologically different leaves in an age-dependent manner, a phenomenon known as heteroblasty. In Arabidopsis thaliana, the SAM produces small orbicular leaves in the juvenile phase, but gives rise to large elliptical leaves in the adult phase. Previous studies have established that a developmental decline of microRNA156 (miR156) is necessary and sufficient to trigger this leaf shape switch, although the underlying mechanism is poorly understood. Here we show that the gradual increase in miR156-targeted SQUAMOSA PROMOTER BINDING PROTEIN-LIKE transcription factors with age promotes cell growth anisotropy in the abaxial epidermis at the base of the leaf blade, evident by the formation of elongated giant cells. Time-lapse imaging and developmental genetics further revealed that the establishment of adult leaf shape is tightly associated with the longitudinal cell expansion of giant cells, accompanied by a prolonged cell proliferation phase in their vicinity. Our results thus provide a plausible cellular mechanism for heteroblasty in Arabidopsis, and contribute to our understanding of anisotropic growth in plants.

A spatiotemporally regulated transcriptional complex underlies heteroblastic development of leaf hairs in Arabidopsis thaliana.

Heteroblasty refers to a phenomenon that a plant produces morphologically or functionally different lateral organs in an age-dependent manner. In the model plant Arabidopsis thaliana, the production of trichomes (epidermal leaf hairs) on the abaxial (lower) side of leaves is a heteroblastic mark for the juvenile-to-adult transition. Here, we show that the heteroblastic development of abaxial trichomes is regulated by a spatiotemporally regulated complex comprising the leaf abaxial fate determinant (KAN1) and the developmental timer (miR172-targeted AP2-like proteins). We provide evidence that a short-distance chromatin loop brings the downstream enhancer element into close association with the promoter elements of GL1, which encodes a MYB transcription factor essential for trichome initiation. During juvenile phase, the KAN1-AP2 repressive complex binds to the downstream sequence of GL1 and represses its expression through chromatin looping. As plants age, the gradual reduction in AP2-like protein levels leads to decreased amount of the KAN1-AP2 complex, thereby licensing GL1 expression and the abaxial trichome initiation. Our results thus reveal a novel molecular mechanism by which a heteroblastic trait is governed by integrating age and leaf polarity cue in plants.

[Semen quality of infertile men carrying hepatitis B virus].

OBJECTIVE: To study the semen parameters of infertile men carrying hepatitis B virus (HBV) and the influence of HBV infection on semen quality. METHODS: We collected the semen samples from 782 infertility males aged 25－35 years old. According to the results of serological examinations, we divided the patients into groups A (HBsAg, HBeAb and HBcAb positive, n = 286), B (HBsAg, HBeAg and HBcAb positive, n = 230) and C (non-HBV control, n = 266), and comparatively analyzed the routine semen parameters, sperm acrosin activity, sperm DNA fragmentation index (DFI) and high DNA stainability (HDS) among the three groups of patients. RESULTS: Compared with the patients of groups B and C, those of group A showed markedly decreased sperm concentration (ï¼»88.20 ± 82.62ï¼½ and ï¼»89.29 ± 53.80ï¼½ vs ï¼»71.49 ± 60.03ï¼½ ×106/ml, P<0.05), progressively motile sperm (PMS) (ï¼»34.88 ± 15.60ï¼½% and ï¼»37.82 ± 13.63ï¼½% vs ï¼»30.70 ± 14.79ï¼½%, P<0.05), sperm motility (ï¼»45.77 ± 16.58ï¼½% and ï¼»48.16 ± 14.03ï¼½% vs ï¼»42.67 ± 17.23ï¼½%, P<0.05), sperm viability (ï¼»82.55 ± 7.55ï¼½% and ï¼»85.26 ± 6.39ï¼½% vs ï¼»81.07 ± 10.19ï¼½%, P>0.05) and morphologically normal sperm (MNS) (ï¼»6.93 ± 4.45ï¼½% and ï¼»7.27 ± 4.43ï¼½% vs ï¼»5.72 ± 3.47ï¼½%, P<0.05), with sperm concentration, PMS, sperm motility, sperm viability and MNS remarkably lower in group B than in C. Sperm acrosin activity was significantly reduced in group A in comparison with groups B and C (ï¼»57.07 ± 26.38ï¼½ vs ï¼»63.03 ± 28.75ï¼½ and ï¼»78.00 ± 33.49ï¼½ µIU/106, P<0.01), remarkably lower in group B than in C (P<0.01). The sperm DFI and HDS, however, were markedly higher in group A than in B (ï¼»14.79 ± 9.46ï¼½% vs ï¼»12.95 ± 7.29ï¼½% and ï¼»11.60 ± 5.98ï¼½%, P<0.05; ï¼»9.62 ± 6.20ï¼½% vs ï¼»8.43 ± 4.72ï¼½% and ï¼»8.41 ± 4.59ï¼½%, P<0.05), and both higher in group B than in C. CONCLUSIONS: Semen quality is lower in infertile men carrying HBV and therefore HBV infection is one of the causes of male infertility.

Telomerase-null survivor screening identifies novel telomere recombination regulators.

Telomeres are protein-DNA structures found at the ends of linear chromosomes and are crucial for genome integrity. Telomeric DNA length is primarily maintained by the enzyme telomerase. Cells lacking telomerase will undergo senescence when telomeres become critically short. In Saccharomyces cerevisiae, a very small percentage of cells lacking telomerase can remain viable by lengthening telomeres via two distinct homologous recombination pathways. These "survivor" cells are classified as either Type I or Type II, with each class of survivor possessing distinct telomeric DNA structures and genetic requirements. To elucidate the regulatory pathways contributing to survivor generation, we knocked out the telomerase RNA gene TLC1 in 280 telomere-length-maintenance (TLM) gene mutants and examined telomere structures in post-senescent survivors. We uncovered new functional roles for 10 genes that affect the emerging ratio of Type I versus Type II survivors and 22 genes that are required for Type II survivor generation. We further verified that Pif1 helicase was required for Type I recombination and that the INO80 chromatin remodeling complex greatly affected the emerging frequency of Type I survivors. Finally, we found the Rad6-mediated ubiquitination pathway and the KEOPS complex were required for Type II recombination. Our data provide an independent line of evidence supporting the idea that these genes play important roles in telomere dynamics.

Phenolic compounds from the twigs and leaves of Tara (Caesalpinia spinosa).

Two new homoisoflavans, 4'-hydroxy-7-methoxy-3-benzyl-2H-chromene (1) and 3,4-cis-di-O-3-hydroxy-7-methoxy-3-(4-hydroxybenzyl)-4-ethoxychroman (2), one new coumarin, 7-methoxy-3-(4-hydroxybenzyl)coumarin (4), together with seven known phenolic compounds, bonducellin (3), anemarcoumarin A (5), (+)-syringaresinol (6), curuilignan D (7), scopoletin (8), and p-hydroxybenzaldehyde (9), were isolated from Tara (Caesalpinia spinosa Kuntze). The structures of the new compounds were characterized from their 1D and 2D NMR spectral data. All the compounds were isolated from this plant for the first time.

[Phenolic compounds from Caesalpinia minax].

Fifteen compounds were obtained from the twigs and leaves of Caesalpinia minax. Their structures were identified as apigenin (1), 5,7,3',4'- tetrahydroxy-3-methoxyflavone (2), luteolin-5, 3 '-dimethyl-ether (3), thevetiaflavon (4), apigenin-7-O-beta-D-glucuronide (5), bonducellin (6), 7-hydroxy-3-( 4-hydroxybenzylidene )-chroman-4-one (7), 3-deoxysappanchalcone (8), 5-acetonyl-7-hydroxy-2-methyl chromone (9), 4-(trans)-acetyl-3,6,8-trihydroxy-3-methyl-dihydronaphthalenone (10), 4-(cis)-acetyl-3,6,8-trihydroxy-3-methyl-dihydronaphthalenone (11), vanillic acid (12), omega-hydroxypropioquaiacone (13), syringaresinol (14) and uracil (15). All compounds were isolated from C. minax for the first time. Compounds 1-14 were phenolic compounds and compounds 1-5, 9-13 and 15 were isolated from the genus Caesalpinia for the first time.

Self-assembled nanoparticles of cholesterol-conjugated carboxymethyl curdlan as a novel carrier of epirubicin.

The purpose of this study was to develop nanoparticles made of cholesterol-conjugated carboxymethyl curdlan (CCMC) entrapping epirubicin (EPB) and establish their in vitro and in vivo potential. CCMC was synthesized and characterized by Fourier transform infrared spectra (FT-IR) and proton nuclear magnetic resonance spectra ((1)H NMR). The degrees of substitution (DS) of the cholesterol moiety were 2.3, 3.5 and 6.4, respectively. EPB-loaded CCMC-3.5 nanoparticles were prepared by the remote loading method. The physicochemical characteristics, drug loading efficiency and drug release kinetics of EPB-loaded CCMC-3.5 nanoparticles were characterized. The in vitro release profiles revealed that EPB release was sensitive to the pH as well as the drug loading contents. The cellular cytotoxicity and cellular uptake were accessed by using human cervical carcinoma (HeLa) cells. The EPB-loaded CCMC-3.5 nanoparticles were found to be more cytotoxic and have a broader distribution within the cells than the free EPB. The in vivo pharmacokinetics and biodistribution were investigated after intravenous injection in rats. Promisingly, a 4.0-fold increase in the mean residence time (MRT), a 4.31-fold increase in the half-life time and a 6.69-fold increase in the area under the curve (AUC 0-->infinity) of EPB were achieved for the EPB-loaded CCMC-3.5 self-assembled nanoparticles compared with the free EPB. The drug level was significantly increased in liver at 24 and 72 h; however, it decreased in heart at 8 and 24 h compared with the free EPB. The in vivo anti-tumor study indicated that the EPB-loaded CCMC-3.5 self-assembled nanoparticles showed greater anti-tumor efficacy than the free EPB. Taken together, the novel CCMC self-assembled nanoparticles might have potential application as anti-cancer drug carriers in a drug delivery system due to good results in vitro and in vivo.

Cationic oxidized microporous rice starch: Preparation, characterization, and properties.

The combination of enzymolysis of compound enzyme, oxidation of sodium hypochlorite, and cationic etherification of 3-chloro-2-hydroxypropyl trimethyl ammonium chloride (CHPTMA) was selected for the functionalization of rice starch (RS) to better raise the performances. The results showed that the oxidation and etherification could improve the acid and alkali resistance of RS, and enhanced its thermal stability. The crystalline structure of RS was an A-type, the enzymolysis, oxidation, and etherification did not change the structural type, while the crystallinity degree of RS derivatives was all reduced. The enzymolysis, oxidation, and etherification altered the pasting properties of RS, and could effectively decrease the setback and breakdown of RS. The oxidation of sodium hypochlorite not only damaged RS particles containing no micropores, but also destroyed the particles containing the micropores. The enzymolysis and oxidation more seriously destroyed the crystalline region than cationic etherification. The oxidation could increase the enthalpy change of RS, whereas the enzymolysis and etherification decreased its enthalpy change. In addition, the enzymolysis and oxidation could lead to the evident increase in average size of RS. The cationic etherification was able to improve the adsorption of Cu2+ on RS, whereas the low oxidation could only slightly ameliorate the adsorption of Cu2+ . PRACTICAL APPLICATION: Cationic oxidized microporous rice starch as an adsorbent, slow-release agent, and flocculant will be well used in food, medicine, pesticide, papermaking, waste water treatment, and so on owing to its abundant micropores, anionic groups, and cationic groups as well as small particle size and narrow size range.

[Different expression of liver-specific insulin-like growth factor I gene in breast cancer; expression with mice].

OBJECTIVE: To investigate the possible relationship between liver-specific insulin-like growth factor I (IGF-1) and the pathogenesis of breast cancer. METHODS: Fifty non-IGF-1 deficient (LID) mice were randomly divided into 2 equal groups: Group I, fed with dimethyl-benzanthracene (DMBA) for 8 weeks to cause mammary cancer, and Group III, fed with DMBA and ginsenoside Rg for 28 d; and 50 LID mice were randomly divided into 2 equal groups too Group II, fed with DMBA for 8 weeks, and Group IV, fed with DMBA and ginsenoside Rg for 28 d. The mice were killed after the cessation of DMBA use. The serum IGF-1 expression was detected with method Six Gene chips were used to detect the gene expression in the breast cancer tissues. RESULTS: The breast cancer rates were 66.67% in Group I and 33.33% in Group II, 36.00% in Group III, and 12.00% in Group IV. The tumor size was (0.79 +/- 0.20) cm in Group I, (0.37 +/- 0.08) cm in Group III , (0.32 +/- 0.08) cm in Group II, and (0.15 +/- 0.05) cm Group IV. The IGF-1 level of Group II was (41.33 +/- 7.52) ng/ml, 1/4 as high as that of Group I [(166.51 +/- 12.32) ng/ml], and the IGF-1 level of Group IV was (33.48 +/- 6.73) ng/ml, 1/4 as high as that of Group III [(155.84 +/- 11.34) ng/ml]. Compared with those of the control mice, the breast cancers of the LID mice had longer latency, lower incidence, and slower growth rate. The differential gene expression in different serum IGF-1 levels involved binding, metabolism, apoptosis, cell cycle, signal transduction, immune response, transcription regulation and interpretation regulation and so on. Among these genes, Col11, Egln3, Glycam1, Irf6, Lgals7, Perp, Rag1, and Rbm35a genes were closely related to the incidence of breast cancer. CONCLUSION: IGF-1 plays a role as a risk factor in the onset and development of breast cancer by affecting the expression of many differentially expressed genes.

[Analysis of apoptosis-related gene expression in different serum level of insulin-like growth factor-1 in mice breast cancer tissue].

OBJECTIVE: A stable primary breast cancer model in liver-specific insulin-like growth factor 1 (IGF-1) deficient (LID) mice and control mice was established. To screen apoptosis related genes expression in different serum IGF-1 levels by gene chip and flow cytometry. METHODS: The LID mice and control mice were used. Induction of breast cancer was achieved by using the 7,12-dimethylbenz(a) anthracene. Ginsenoside Rg3 was used to interfering therapy treatment. The incidence of breast cancer in every group was compared, and expression of apoptosis associated genes was detected by gene chip and flow cytometry. RESULTS: The incidence of tumor in none ginsenoside Rg3 injected control mice was 66.7%. The incidence of tumor in ginsenoside Rg3 injected LID mice was 12.0% which was significantly lower than any other group (P < 0.05). The apoptosis percentage in none ginsenoside Rg3 injected control mice was (2.7 +/- 0.7)%. The apoptosis percentage in ginsenoside Rg3 injected LID mice was (14.0 +/- 1.7)%. The results of gene chip indicated that in contrast to LID mice, LTA, LTB, TNF-alpha, TRAIL, TRANCE, BLK, BOK, CASP8, TRAF5, and APAF1 genes were down-regulated, and LTBR, TRAF4 genes were up-regulated in the breast cancer tissues of control mice. Application of ginsenoside Rg3 therapy could change the expression of these genes. CONCLUSIONS: Circulating IGF-1 levels play a role in the onset and development of breast cancer. Degrade serum IGF-1 level is able to promote apoptosis by affecting the expression of a series of apoptosis related genes consequently inhibit the growth of breast cancer. There was a synergistic effect with the application of ginsenoside Rg3.

[Light-dependent ethylene signal abnormality of the arabidopsis auxin-related uro mutant].

The Arabidopsis semi-dominant mutant upright rosette (uro) was named from its upright-growing leaves. Previous results suggest that uro mutant was caused by a single gene mutation and the URO function might be associated with auxin-mediated plant development. In this paper, some ethylene-related phenotypes of uro mutant are reported, such as hyponastic rosette leaves, persistent off perianth and elongated hypocotyls. Results of ACC and ethylene inhibitor AgNO(3) treatment experiment suggest that some of the phenotypes of uro mutant are caused by ethylene signal system abnormality. uro mutant is an important material for studying the ethylene signal system especially for the relationship between auxin and ethylene.

Inflammation and airway remodeling of the lung in guinea pigs with allergic rhinitis.

Allergic rhinitis (AR) and asthma belong to the category of type I allergic diseases, whose pathological features are airway remodeling of the lung and allergic inflammation. The aim of the present study was to evaluate inflammation and remodeling of lung tissue in a guinea pig model of AR in order to confirm consistent pathological changes of upper and lower airways in AR. Male guinea pigs were randomly divided into an experimental and a control group (n=10 in each). The AR model was established by sensitization through intraperitoneal injection of ovalbumin for three weeks and bilateral nasal local excitation for twelve weeks. All tissues of nasal mucosa and lung were subjected to hematoxylin and eosin as well as toluidine blue staining, and characteristics of remodeling of lung tissue, including thickness of bronchial wall, epithelial mucosa and smooth muscle were histologically determined. Collagen deposition in lung tissue was observed by Masson's trichrome stain. Severe paroxysmal nose scratching action, frequent sneezing, visible outflow of secretion from the anterior naris and frequent nose friction were observed in the AR model group within 30 min after local excitation. The total symptom scores were significantly increased in the AR model group compared with those in the control group. Obvious inflammatory cell infiltration was observed in the AR model group. Compared with those in the control group, the numbers of eosinophils and mast cells in nasal mucosa and lung tissue were significantly increased. Obvious airway remodeling of the lung was observed in the AR model group. Compared with those in the control group, bronchial wall thickness, epithelial layer thickness and smooth muscle thickness in the airways were significantly increased in the AR model group. Increased collagen deposition was found in the AR model group compared with that in the control group. The results of the present study revealed that inflammation and airway remodeling of lungs arose in guinea pigs with AR, suggesting that pathological changes of upper and lower airways are consistent in this AR model.

Genetic association between TNF-α -857 C/T polymorphism and ankylosing spondylitis susceptibility: evidence from a meta-analysis.

Certain studies have suggested that the tumor necrosis factor-α (TNF-α) -857 C/T polymorphism is associated with risk of ankylosing spondylitis. However, the conclusions remain controversial. Therefore, we performed a meta-analysis to provide a more precise conclusion. Such databases as PubMed, Embase, CBM, CNKI, and Wanfang Data were searched to identify relevant studies up to August 26, 2015. Odds ratios (ORs) and 95% confidence intervals (CIs) were used to estimate the association between TNF-α -857 C/T polymorphism and ankylosing spondylitis susceptibility. A total of 10 studies were included in the meta-analysis. Overall, an elevated risk between TNF-α -857 C/T polymorphism and ankylosing spondylitis was observed in three genetic model (T vs. C: OR 1.86, 95% CI 1.19-2.92; CT vs. CC: OR 2.51, 95% CI 1.49-4.23; TT + CT vs. CC: OR 2.46, 95% CI 1.40-4.30), except in homozygote model (TT vs. CC: OR 2.41, 95% CI 0.96-6.06) and recessive model (TT vs. CT + CC: OR 1.54, 95% CI 0.71-3.35). Sensitivity analysis showed the overall results were robust. Subgroup analyses according to Hardy-Weinberg equilibrium and ethnicity showed that the increased risk of ankylosing spondylitis were predominant in Asian population. This meta-analysis indicated that TNF-α -857 C/T polymorphism might increase the susceptibility of ankylosing spondylitis, especially in Asians. Further studies were needed to verify the conclusion.

[Pollution Characteristics and Risk Assessment of Polybrominated Diphenyl Ethers in the Surface Sediments from Middle Reaches of the Yangtze River].

Polybrominated diphenyl ethers (PBDEs) are highly lipophilic compounds with high toxicity and long-term bioaccumulation, which have strong affinities to suspended particulates in water. So PBDEs are easily adsorbed to sediments, and cause potential risks to human beings and aquatic ecosystem. In order to investigate the levels and patterns of PBDEs in the Yangtze River, 13 surface sediment samples were collected from the middle reaches of the regions and the concentrations of 9 PBDE congeners in the sediments were determined using High Resolution Gas Chromatography/High Resolution Mass Spectrometry (HRGC/HRMS). The total concentrations of PBDEs ranged from 46.1 to 326 pg · g⁻¹ dry weight. BDE-99 and BDE-47 were the dominant PBDE congeners, which averagely contributed 51.6% and 19.6%, respectively. The concentrations of PBDEs were relatively low compared to those in other studies. The total organic carbon (TOC) was also analyzed in this study, and the results indicated that there was no obvious positive correlation between the levels of PBDEs and TOC. In addition, based on the PBDEs concentration levels, the ecotoxicological risks of PBDEs contamination in surface sediments of the Yangtze River were evaluated by the quotient method. The results showed that the levels of PBDEs were considerably low and these compounds should have no risk to human health.

[Candida infection in patients with acute necrotizing pancreatitis].

OBJECTIVE: To summarize our hospital's experience in the diagnosis and treatment of Candida infection in patients with acute necrotizing pancreatitis (ANP). METHODS: Seventy-eight cases with ANP were reviewed. There were diagnoses either by operative finding or by CT scanning. Sixty-two cases received prophylactic antibiotic treatment, other sixteen did not. For cultivation of Candida, blood, urine, stool, sputum and wound drainage fluid culture, and swabs were examined microbiologically for fungi. RESULTS: The incidence of Candida infection in all patients with ANP was 17.9% (14/78) and mortality was 28.6% (4/14). The incidence of prophylactic antibiotic group was 19.4% (12/62) and mortality was 25.0% (3/12). Non prophylactic group was 12.5% (2/16) and 50.0%. CONCLUSIONS: Our data provide evidence for the clinical significance of Candida infection in patients with ANP. The current prophylactic antibiotic treatment can prevent a septic course of the ANP, but might lead to the evolution of Candida infection.

Stability and in vivo evaluation of pullulan acetate as a drug nanocarrier.

To develop pullulan acetate nanoparticles (PANs) as a drug nanocarrier, pullulan acetate (PA) was synthesized and characterized. Its acetylation degree determined by the proton nuclear magnetic resonance ((1)H NMR) was 2.6. PANs were prepared by the solvent diffusion method and characterized by transmission electron microscope (TEM), size distribution, and zeta potential techniques. PANs had nearly spherical shape with a size range of 200-450 nm and low zeta potentials both in distilled water and in 10% FBS. The storage stability of PANs was observed in distilled water. PANs were stored for at least 2 months with no significant size and zeta potential changes. The safety of PANs was studied through single dose toxicity test in mice, and the result showed that PANs were well tolerated at the dose of 200 mg/kg in mice. Epirubicin-loaded PANs (PA/EPI) were also prepared and characterized in this study. Moreover, the in vivo pharmacokinetics of PA/EPI was investigated. Compared with the free EPI group, the PA/EPI group exhibited higher plasma drug concentration, longer half-life time (t(1/2)) and the larger area under the curve (AUC). All results suggested that PANs were stable, safe, and showed a promising potential on improving the bioavailability of the loaded drug of the encapsulated drug.

[Correlation of insulin-like growth factor-1 (IGF-1) to angiogenesis of breast cancer in IGF-1-deficient mice].

BACKGROUND & OBJECTIVE: Insulin-like growth factors (IGFs) play important roles in the development and progression of tumors. But the mechanism of tumorigenesis in relation to IGF-1 is unclear yet. This study was to explore the correlation of circulating IGF-1 level to the angiogenesis of breast cancer in IGF-1-deficient mice. METHODS: The liver-specific IGF-1-deficient (LID) mice and control mice were injected with 7,12-dimethybenz(a)anthracene (DMBA) to develop breast cancer. Ginsenoside Rg3 was used to intervene tumor growth. The occurrence rates of breast cancer were compared. The expression of vascular endothelial growth factor (VEGF) and microvessel density (MVD) was detected by immunohistochemistry. RESULTS: The occurrence rate of breast cancer was 66.67% in untreated control mice, 33.33% in untreated LID mice, 36.00% in Rg3-treated control mice, and 12.00% in Rg3-treated LID mice. The tumor size was (0.79+/-0.20) cm in untreated control mice, (0.37+/-0.08) cm in untreated LID mice, (0.32+/-0.08) cm in Rg3-treated control mice, and (0.15+/-0.05) cm in Rg3-treated LID mice. The average light density and positive rate of VEGF were the highest in untreated control mice (0.34+/-0.10 and 0.04+/-0.02, P<0.05), and the lowest in Rg3-treated LID mice (0.13+/-0.03 and 0.01+/-0.00, P<0.05). The MVD was 31.9+/-5.3 in untreated control mice, 26.8+/-4.9 in untreated LID mice, 20.1+/-4.9 in Rg3-treated control mice, and 14.4+/-4.9 in Rg3-treated LID mice. CONCLUSIONS: Circulating IGF-1 plays a role in the onset and development of breast cancer. Degrading serum IGF-1 level could inhibit angiogenesis and growth of breast cancer. Rg3 could promote this effect.

Genistein stimulates the osteoblastic differentiation via NO/cGMP in bone marrow culture.

The soybean phytoestrogen, genistein (Gen), has anabolic effects on bone through mechanisms that remain to be elucidated. We examined the role of nitric oxide (NO) and its downstream effector guanylyl cyclase (GC) in mediating the effects of Gen on the proliferation and osteoblastic maturation of primary mouse bone marrow-derived mesenchymal stem cells (BMSCs). Gen (10(-8) approximately 10(-6) M) resulted in a dose-dependent increase in cell proliferation as measured by increased [3H]thymidine incorporation, and stimulated osteoblastic maturation as assessed by culture duration-dependent increments in alkaline phosphatase (ALP) activity, calcium deposition into extracellular matrix and Runx2/Cbfa1 gene expression in BMSCs cultures. Gen also resulted in a dose-dependent increase in NO synthase (NOS) activity, NO formation, and cGMP production in BMSCs cultures. The effects of Gen were mimicked by 17beta-estradiol (E2, 10(-8) M). Concurrent treatment with the estrogen receptor (ER) antagonist ICI182,780 (10(-7) M) or the NOS inhibitor L-NAME (3 x 10(-3) M) diminished the Gen (10(-6) M)-mediated increase in NOS activity, NO production, and cGMP content. In contrast, a soluble GC inhibitor 1H-[1,2,4]oxadiazolo [4,3,-a]quinoxalin-1-one (ODQ, 10(-6) M) selectively blocked the Gen (10(-6) M)-mediated increase in cGMP content but not in NO production and NOS activity. Moreover, inhibition of ER, NOS activity or cGMP blocked Gen-induced proliferation and osteoblastic differentiation of BMSCs and Runx2/Cbfa1 gene expression in culture. Gen has estrogen-like activity and stimulates the proliferation and osteoblastic differentiation of mouse BMSCs at least in part through NO/cGMP pathway.