Effects of PD98059 on the differentiation from mesenchymal stem cells to osteoblasts

AIM: To investigate the effects of PD98059 on the differentiation from mesenchymal stem cells to osteoblasts.METHODS: hMSC were separated from human marrow and expanded in cuture medium. hMSC were induced with dexamethasone, β-glycerophosphate, vitamin C which acted as osteoblast differentiation inducer. PD98059 was added into the osteoblasts induction medium. The cells were assayed with cell morphology, alkaline phosphatase (AP) activity and calcium deposition. RESULTS: The isolated cultured MSC comprised a single phenotypic population and displayed a fibroblast-like morphology. After induced with osteoblasts induction medium, the cells showed changes in cell morphology from spindle-shape to cuboidal and polygonal. The AP activity increased gradually and reached the peak in 12 days, then decreased. Many scattered tangerice calcium nodes were observed. PD 98059 significantly inhibited AP activity and calcium deposition in a dose-dependent manner. A striking observation of the present study was that a few adipocytes appeared in cultures that were treated with PD 98059 and osteogenic differentiation medium. CONCLUSION: These results indicated that osteogenic diferentiation from the hMSCs was related to the activation of the ERK.     

B allele in Iα1 hs1,2 VNTR region is associated with IgA nephropathy

AIM: To investigate the relationships between Iα1 hs1,2 VNTR polymorphism and IgA nephropathy. METHODS: Four hundred and ninteen patients with IgA nephropathy and their first-degree relatives were recruited. Two hundred and one sex and age-matched normal Chinese Han volunteers were also recruited as controls. After extracting genomic DNA, the VNTR genotypes of Iα1 hs1,2 region were determined by PCR and electrophoresis, and the results were analyzed by transmission disequilibrium test (TDT) and haplotype relative risk (HRR) in the families, and Chi-Square test in the case-control analysis. RESULTS: ① TDT analyses showed that B allele of the Iα1 hs1,2 VNTR region was significantly more transmitted from heterozygous parents to patients than expected (101 Trios, χ2=6.818, P＜0.01), extended TDT produced the same results (164 families, χ2=7.583, P＜0.01). ② Consistent with the TDT results, HRR also showed that B allele was over-transmitted to patients (P＜0.05, χ2=4.122, HRR=1.180), and the BB genotype conferred a higher risk of developing the disease (P＜0.05, χ2=4.411, OR=1.538). ③ The case-control study indicated that the B allele had a higher frequency in the IgA nephropathy group (χ2=6.968, P＜0.05). CONCLUSION: B allele in Iα1 hs1,2 VNTR region is associated with susceptibility to IgA nephropathy.     

Effect of rhIL-10 on IL-6 and TNF-α levels in serum and liver of lipopolysaccharide-challenged mice

AIM: To investigate the effect of recombinant human interleukin-10 (rhIL-10) on IL-6 and TNF-α levels in serum and liver of mice exposed to lipopolysaccharide (LPS). METHODS: rhIL-10 was prepared by using genetic engineering technology. Mice were intraperitoneally with 500 μg of LPS, and then were treated intravenously with various dosages of rhIL-10. The levels of IL-6 and TNF-α in hepatic tissue and serum were determined by ELISA at 12 h, 24 h, 48 h and 72 h post rhIL-10 treatment. RESULTS: rhIL-10 markedly inhibited the increase in IL-6 and TNF-α levels in hepatic tissue and serum at 12 h after rhIL-10 treatment in LPS-challenged mice, and the inhibition effect was significant at 24-48 h after rhIL-10 treatment （P＜0.05）. CONCLUSION: rhIL-10 can inhibit the increase in IL-6 and TNF-α levels induced by LPS in mice.     

Preparation and identification of anti- angiostatin monoclonal antibodies

AIM:To further study the anti-tumor effect of angiostatin, an anti-human angiostatin monoclonal antibody was prepared and identified.METHODS:The hybrodoma techniques were used. The BALB/C mice were immunized with angiostatin. The supernatant of cell culture were collected and screened by ELISA and double immunodiffusion.RESULTS:There cell lines which steadily secreted the anti-angiostatin monoclonal antibody were identified by ELISA and double immunodiffusion. The antibody was IgG1 and specifically recognized angiostatin without crossing reactions to rhIL-2, rhTNF-α, rhIFN-α and serum proteins.CONCLUSION:The antibodies secreted by three hybridoma cell lines identified by several methods were specific antibodies of angiostatin.     

Adult human mesenchymal stem cell differentiates into adipocytes

AIM:To investigate the differentiation from human mesenchymal stem cells (hMSC) to adipocytes.METHODS: hMSC were separated from rib marrow and expanded in culture medium. To detect the surface antigens, the labeled cells were analysed on a FACScan flow cytometer. hMSC were induced with dexamethasone, insulin, 1-methy1-3-isobutylxanthine and indomethacin which acted as adipocyte differentiation inducer. The cells were stained with Oil Red O. The number of adipocytes were counted on a phase-contrast microscope.RESULTS: hMSC were expanded as undifferentiated cells in culture for more than 5 passages. The isolated cultured MSC comprised a single phenotypic population and displayed a fibroblast-like morphology. These expanded, attached MSC were uniformly positive for CD29, CD44, CD90, CD105, CD166 and didn't express CD14, CD34, CD45, CD11a. After induced with induction medium, lipid vacuoles were first detectable within the cells at 48 hours. Two weeks later, more than 85% MSC differentiated into adipocytes which displayed a perinuclear accumlation of lipid vacuoles, as detected by Oil Red O. CONCLUSION:hMSC can be induced to differentiate into adipocytes.     

葡萄果实发育后期脱落酸来源的研究

  以‘品丽珠’(Vitis vinifera L. cv. Cabernet-Franc) 葡萄为试材, 对发育后期果实中脱落酸(ABA) 的来源进行了研究。结果表明, 叶片合成的ABA 可能是果实中ABA 的重要来源, 而根部ABA 的合成对果实中ABA 含量贡献不大。种子中ABA 的合成可能不是果肉中ABA 的重要来源。果实具有独立合成和降解ABA 的能力。对果实内ABA 含量变化的代谢调控问题进行了讨论。     

Human mesenchymal stem cells differentiate into neuron-like cells with Tanshinone II A

AIM:To investigate the differentiation from human mesenchymal stem cells(hMSC) into neuron-like cells with Tanshinone II A.METHODS:hMSC were separated from rib marrow with Ficoll-Paque reagent and expanded in culture medium. To detect the surface antigens, the labeled cells were analysed on a FACScan flow cytometer to determine the effect of the capacity of proliferation and differentiation of the mesenchymal stem cells with FGF-2. hMSC were induced to differentiate into neurons with DMEM Tanshinone II A. Neuron-specific enolase(NSE), neurofilament(NF), Nestin, glial fibrillary acaidic protein(GFAP) were detected by immunohistochemistry.RESULTS:hMSC were expanded as undifferentiated cells in culture for more than 15 passages. The isolated cultured MSC comprised a single phenotypic population and displayed a fibroblast-like morphology. These expanded attached MSC were uniformly positive for CD29, CD44, CD90, CD105, CD166 and didn't express CD11a, CD14, CD34, CD38, CD45, CD80, CD86. FGF-2 have special effect on low denisity MSCs. Simple methods with Tanshinone II A induced hMSC to exhibit a neuronal phenotype, expressing NSE, NF-M, Nestin at 5 hours. But the neuron-like cells didn't express the glial astrocyte marker GFAP.CONCLUSION:hMSC can be induced to differentiate into neurons with Tanshinone II A.     

我国新药研发中专利文献的利用策略

分析了我国新药研究开发在专利文献利用过程中存在的问题,探讨了合理利用专利文献进行新药研究与开发的6项策略,包括重视专利文献检索,加强对专利法律状态的分析与研究,利用同族专利了解新药的技术水平与经济价值,调查专利申请数量和预测新药研发趋势,充分利用专利权的限制性条款,与加快培养高素质的专利信息人才。

     

Fuzi polysaccharide protects neonatal rat cardiomyocytes with hypoxia-reoxygenation by inhibiting endoplasmic reticulum stress

AIM: To investigate whether the protection mechanism of Fuzi polysaccharide (FPS) is related to inhibition of endoplasmic reticulum stress in cultured neonatal rat cardiomyocytes with hypoxia/reoxygenation (H/R). METHODS: Cultured rat myocardial cells were divided into control group, H/R group (hypoxia for 3 h and reoxygenation for 6 h) and different concentrations of FPS (0.1 g/L, 1 g/L, 10 g/L or 20 g/L) +H/R groups. The cell survival was detected by MTT assay and cell apoptosis of cardiomyocytes was measured by flow cytometry using Annexin V-FITC staining. The expression of glucose-regulated protein 78 (GRP78), CCAAT/enhancer-binding protein homologous protein (CHOP) and caspase-12 were determined by Western blotting. The mRNA expression of CHOP and caspase-12 was detected by quantitative PCR. RESULTS: After reoxygenation, the expression of GRP78, CHOP and caspase-12 in cardiomyocytes was increased. Compared with H/R group, the expression of GRP78, CHOP and caspase-12 in FPS+H/R groups was significantly inhibited, the survival rate of cardiomyocytes was increased and the apoptosis of cardiomyocytes was inhibited. This protective effect of FPS was in a dose-dependent manner and reached its peak at 10 g/L. CONCLUSION: Fuzi polysaccharide protects cardiomyocytes from H/R injury. The mechanism is related to inhibiting endoplasmic reticulum stress.     

Pretreatment with Sini decoction induces delayed preconditioning in rat heart and role of p38 MAP kinase

AIM: The present study was designed to determine whether Sini decoction (SND), a traditional Chinese medicine, induces delayed preconditioning-like effect in rat heart and the possible mechanism by which ischemia myocardium is protected. METHODS: Sprage－Dawleyt rats underwent three 5 min episodes of preconditioning ischemia 24 h prior to the global ischemia and reperfusion in ischemic preconditioning/second window of protection (IPC/SWOP) group or were pretreated with Sini decoction (5 mL·kg-1·d-1 for 3 days, the last treatment 24 h before global ischemia and reperfusion) in SND group. Myocardial infarct size, CK, LDH and NO were examined. p38 MAPK and PKC were determined by immunohistochemisty. RESULTS: Myocardial infarct size was significantly decreased, CK and LDH were decreased in the serum, NO2-/NO3- was increased in myocardial tissue in SND group as well as in IPC/SWOP group (there was no difference between the two groups). The expression of p38 MAPK and PKC were upregulated in myocardial tissue in SND and IPC/SWOP groups. CONCLUSION: These results suggest that Sini decoction induces delayed preconditioning-like effect in the rat heart, possibly via inducing p38 MAPK activation.