Study on growth and metabolites accumulation of Ganoderma lucidum mycelium by cadmium stress / 中草药

Objective: To study the effects of cadmium stress on mycelial growth and accumulation of metabolites in Ganoderma lucidum, and to explore the mechanisms affecting growth and accumulation of metabolites, and to provide evidence for controlling cadmium in the production and cultivation of G. lucidum. Methods: The mycelium of G. lucidum was cultured under the conditions of heavy metal ion cadmium concentration of 0, 0.5, 1, 4, 10, and 40 mg/L, and its biomass accumulation, intracellular ROS level, membrane oxidative damage, anti-oxidant enzyme activity, and ROS regulation related enzyme expression were analyzed. Results: When the concentration of cadmium reached 4 mg/L, the mycelial growth was inhibited. The levels of intracellular ROS, H2O2, and MDA increased significantly, increasing by 76%, 46% and 325%, respectively, and increased with the increase of cadmium concentration; The NADPH expression levels of oxidase gene (NOXA), superoxide dismutase gene (SOD1 and SOD4), and CATalase gene (CAT) were significantly up-regulated. When the cadmium concentration reached 10 mg/L, the inhibitory effect was significant. The colony growth diameter and the dry weight inhibition rate of fermentation mycelium were 26.15% and 32.78%, respectively. The total triterpenoid inhibition rate of G. lucidum was 33.7%, and the inhibition rate of total protein synthesis was 30.3%. Inhibition of polysaccharides was not significant. When the cadmium concentration reached 40 mg/L, the expression levels of Ascorbate peroxidase gene (APX) and Glutathione peroxidase gene (GPX) were significantly up-regulated. With the increase of cadmium concentration, the activities of SOD, CAT, APX, and GPX increased first and then decreased. When the concentration of cadmium reached 1 mg/L, the activity of GPX decreased and the activity of APX increased significantly. Exogenous addition of diphenyleneiodonium chloride (DPI), N-acetyl-L-cysteine (NAC) and vitamin C (VC) had significant effects on cadmium-induced G. lucidum clearance of ROS and reduction of MDA content. Conclusion: Cadmium stress causes the decrease of mycelial production and metabolite accumulation of G. lucidum, which may be due to the inhibition of GPX activity by cadmium ions, resulting in the accumulation of H2O2, causing the increase of ROS level and membrane oxidative damage, inhibiting mycelial growth and accumulation of metabolites, and regulating NOX. Up-regulation of gene expression results in an increase in anti-oxidant enzyme activity and expression to increase the clearance of reactive oxygen species. Therefore, the cadmium content should be controlled within the range of 1 mg/L during the production process.

Low dose-intensity docetaxel monotherapy for patients with anthracycline-resistant metastatic breast cancer in poor physical status / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To evaluate the efficacy and safety of a weekly schedule of low dose-intensity docetaxel monochemotherapy for patients with anthracycline-resistant metastatic breast cancer (MBC) in poor physical status.</p><p><b>METHODS</b>Thirty MBC patients who were previously exposed to anthracycline treatment received docetaxel alone at a dose of 30 mg/m2 on D1, D8 and D15, repeated every 4 weeks for a maximum of 6 cycles.</p><p><b>RESULTS</b>Of the 30 evaluable patients, 2 (6.7%) achieved a complete response, and 9 (30.0%) a partial response, with an overall objective response rate of 36.7% (95% CI: 20.5%-53.9%). The most common adverse event was hematologic toxicity. After an average follow-up of 15.0 months, the median time to progression (TTP) was 8. 5 months and the median overall survival (OS) had not reached yet at the end of follow-up.</p><p><b>CONCLUSION</b>The weekly low dose-intensity docetaxel monochemotherapy is effective and well-tolerated in patients with anthracycline-resistant metastatic breast cancer in poor physical status.</p>

Relationship between endostatin and vascular cell adhesion molecule-1 expressions on bone marrow stromal cells in BMT-mice / 中国实验血液学杂志

This study was aimed to investigate the relationship between endostatin and vascular cell adhesion molecule-1 (VCAM-1) expressions on bone marrow stromal cells (BMSC) in mice after bone marrow transplantation (BMT) and effect of ligustrazine on their expressions. The mice were randomly divided into 3 groups: normal group (without treatment), saline group (control of BMT) and ligustrazine group (BMT + ligustrazine). BMT mouse models were established. The normal group was not treated, the saline group was given normal saline (0.2 ml/mouse, twice a day) through gastric tube, while the ligustrazine group was given ligustrazine (0.2 ml/mouse, twice a day) also through gastric tube. On day 7, 14, 21 and 28 after BMT, mice were killed by euthanasia. The expression levels of endostatin and VCAM-1 in bone marrow stromal cells were detected by immunohistochemistry and RT-PCR analysis respectively. The results showed that the endostatin protein mainly expressed in nuclei of BMSCs, the VCAM-1 protein mainly expressed in plasma of BMSCs. On day 7, 14, 21 after BMT the expression levels of endostatin mRNA and protein in ligustrazine and saline groups were significantly lower than that in normal group (P < 0.01 or P < 0.05), while their expression levels in ligustrazine group were lower than that in saline group. On day 28 the expression levels in saline group returned to normal, while the expression levels in ligustrazine group not were normalized. On day 7, 14, 21 after BMT the expression levels of VCAM-1 mRNA and protein in ligustrazine and saline groups were significantly lower than that in normal group (P < 0.01 or P < 0.05), but their expression levels in ligustrazine group were significantly lighter than that in saline group (P < 0.01 or P < 0.05). On day 28 the VCAM-1 expression level in ligustrazine group returned to normal, while its expression level in saline group not were normalized. The difference between these two groups was significant (P < 0.01). Correlation analysis revealed that there was a negative correlation between endostatin and VCAM-1 expression in saline group, there was a positive correlation between endostatin and VCAM-1 expression in ligustrazine group. It is concluded that the endostatin can influence hematopoiesis in bone marrow by affecting VCAM-1 expression on BMSC and hindering connection between stromal cells and hematopoietic cells as well as extracellular stroma and hematopoietic cells, while ligustrazine can enhance the adhesion molecule expression on stromal cell surface of bone marrow in BMT-mice, accelerate the homing and proliferation of HSPC in bone marrow after BMT, meanwhile can promote the repair of bone marrow microenvironment, accelerate hematopoietic reconstitution of bone marrow after BMT through feedback regulation of endostatin expression of BMSC in BMT-mice.

Effect of ligustrazine on the expression of bFGF in bone marrow stromal cells of mice after BMT / 中国实验血液学杂志

This study was purposed to investigate the effect of ligustrazine on the expression of bFGF in bone marrow stromal cells (BMSC) and to explore the mechanism of hematopoietic reconstitution after bone marrow transplantation (BMT). The mice were randomly divided into 3 groups: normal group, saline group and ligustrazine group. BMT mouse models were established. The mice of normal group were not treated, the mice of saline group were given normal saline (0.2 ml/mouse, twice a day) through gastric tube, while the mice of ligustrazine group were given ligustrazine (0.2 ml/mouse, twice a day) through gastric tube. On day 7, 14, 21 and 28 after BMT, the femora were taken and the bone marrow mononuclear cell (BMMNC) suspensions were used for the cultivation of bone marrow stromal cells according to Dexter's culture method. The mRNA and protein expressions of bFGF in BMSC were assayed by RT-PCR and Western blot respectively. The results showed that the expression of bFGF in BMSC on the level of mRNA and protein were all reduced significantly after BMT, and increased slowly with the time. On day 7, 14 and 21 after BMT, the expressions of bFGF mRNA and protein in bone marrow stromal cells of ligustrazine group and saline group were lower than that in bone marrow stromal cells of normal group, but the expressions of bFGF mRNA and protein in ligustrazine group were obviously higher than that in saline group (P < 0.01 or P < 0.05). On day 28 after BMT, the expressions of bFGF mRNA and protein in ligustrazine group returned to normal level, while the expressions of bFGF mRNA and protein in saline group not returned to normal level, there was significant difference between these two groups. It is concluded that ligustrazine can enhance bFGF expression level in bone marrow stromal cells after syngeneic bone marrow transplantation in mice, which confirms that ligustrazine can enhance the repair of bone marrow microvessels, improve bone marrow microenvironment and promote hematopoietic reconstitution.