梨果实贮藏中可溶性果胶和半纤维素分子结构的变化

 为了了解分子结构变化与耐藏性的关系，以耐贮性不同的苹果梨和新高梨为试材，利用Ultrogel AcA 22凝胶柱渗漏分离，比较了梨贮藏中可溶性果胶和半纤维素分子大小的变化。结果表明，苹果梨的半纤维素分子大于新高梨。在贮藏过程中，耐贮性强的苹果梨果实可溶性果胶分子大小变化不明显，但半纤维素分子明显变小。相反，新高梨的可溶性果胶分子明显变小，而未观察到半纤维素分子大小的变化。说明梨的耐贮性与可溶性果胶分子结构变化及半纤维素分子大小有关，而与半纤维素分子结构变化没有直接关系。     

Classification of Morchella species from Yunnan and Zhejiang Provinces Based on rDNA ITS Sequences

Fourteen Morchella samples (eleven from Yunnan and three from Zhejiang Provinces) were selected on the basis of differences in fruit body morphology. Ribosomal DNA internal transcribed spacers (ITS) were amplified in each case using the universal primer pair, ITS-1 and ITS-4, and the amplification products were purified and sequenced. Comparisons with sequence data in GenBank revealed that the 11 Morchella isolates collected from Yunnan belonged to four species: Morchella elata, Morchella conica, Morchella crassipes and Morchella costata. The three isolates collected from Zhejiang Province (M12, M13 and M14) were designated as unknown Morchella species. When Verpa conica (AJ544206) (from the genus Verpa belonging to the same family as Morchella) was taken as the outgroup, the 14 isolates formed three groups, M. elata, M. costata (Group 1); Morchella esculenta, M. conica (Group 2); and M. crassipes, M12, M13 and M14 (Group 3).     

切花菊14个品种的核形态学研究

 采用常规压片法对不同花序类型、不同花色的14个切花菊品种进行了核形态学研究。结果表明：（1）间期核均为复杂染色中心型，前期染色体属于中间型。(2)体细胞染色体在品种内和品种间均呈现多数目性，染色体数变动范围在48~56之间，以54条为主。（3）核型呈现多样性，由中部、近中部和近端部着丝粒染色体组成，‘神马’、‘小红娘’、‘九月黄’和‘小菊双色’有1或2条随体染色体；染色体相对长度为2.04%~5.26%；核型不对称系数为62.39%~67.32%；‘长紫’、‘意大利红’、‘小红娘’和‘阿里格斯’核型类型为“2B”，其它品种为“2A”型。（4）有丝分裂后期和末期除‘清露’外均正常。基于观察结果分析了切花菊核型多样性及进化趋势。     

Biological characteristics of JAK2 transduced CD34+ cells from cord blood during ex vivo expansion

AIM: To explore the feasibility and biological characterization of long-term regulated expansion of JAK2 transduced human CD34⁺ cord blood cells in vitro.METHODS: A retrovirus (RV) vector which contains JAK2 catalytic domain and two binding sites for a chemical inducer,dimerization (AP20187),was cloned (designated MGI-F₂JAK2).CD34⁺cells were enriched from cord blood with a MiniMACS system.The purified CD34⁺ cells were transfected with supernatant from the retrovirus packaging cell line that expressed JAK2.Following transduction,cells were expanded into four groups: AP20187 alone,FL alone,TPO,alone,AP20187+FL+TPO,respectively.The expanded cells were monitored by GFP expression,immunophenotyping,progenitor colony assay,karyotype analysis as well as tumorigenesis in nude mice.RESULTS: The purity of selected CD34⁺ cells was over 91% and gene transfer rate was 49.32%±6.21%.Only the group of AP20187 +FL+ TPO was obtained a significant sustained outgrowth of the transduced CD34⁺ cord blood cells.The percentage of GFP+ cells consistently produced a rise to the 90% peak level by the end of 8th week of culture.Flow cytometry analysis showed that the phenotype of the expanded cells was CD33⁺,CD61⁺ and Gly-A⁺ partial positive;CD38⁺ and HLA-DR⁺ strong positive,while CD2,CD7 and CD19 were almost negative.Colony assays performed in methycelluos,which can give rise to BFU-E,CFU-GM and CFU-Mix,the CFU-GM was predominantly in all colonies.The tumor was not observed in nude mice and the karyotype analysis was normal from expanded cells.CONCLUSION: The results demonstrate that AP20187-mediated activation of JAK2 signaling is capable of stimulating expansion JAK2 transduced CB CD34⁺ cells in combination with FL and TPO.This system may have applications for studies in signaling transduction,hematopoiesis,and for gene and cell therapy.     

Effects of cinnamyl aldehyde on c-Fos and c-Myc expression in NIH3T3 cells

AIM: Cinnamyl aldehyde (CA) is one alcohol ingredient derived from Cinnamomum cassia,which is widely used in treating chronic skin wound in Chinese medicine with the curative effect of ‘rescuing YANG’.The purpose of the present study was to investigate the expression of c-Fos,c-Myc proteins at different time points in NIH3T3 treated with CA and explore the possible mechanism of promoting cell proliferation by CA.METHODS: MTT assay was used for observing cell proliferation.Expression of c-Fos and c-Myc proteins in NIH3T3 cells were assessed by immunocytochemistry assay.RESULTS: The cell proliferation was promoted obviously when CA concentration was between 8.8×10^-2 μg/L and 8.8×10 μg/L.CA at concentration of 5.5 μg/L significantly induced expression of c-Fos,c-Myc proteins at 2-3 h after the stimulation compared with control group (P<0.01).CONCLUSION: CA increases expression of c-Fos and c-Myc proteins,which may be one of mechanisms for CA to promote NIH3T3 cell proliferation.     

GM-CSF induces human vascular endothelial cells to form vessel-like structure and the role of VEGF

AIM: To investigate the influence of GM-CSF on human vascular endothelial cells induced to form new blood vessels and the role of VEGF. METHODS: HUVECs were cultured by Matrigel to set up a stable angiogenesis system with the stimulating factors. The rhGM-CSF concentration-dependent and time-dependent effects and the role of VEGF165 were detected. CD34 was measured by immunochemical staining and numbers of vessel formation was calculated under microscopic observation. RESULTS: After treatment with rhGM-CSF at various concentrations and at different time points, the numbers of vessel formation increased in a dose-dependent and time-dependent manner. In the presence of VEGF165, the numbers of vessel formation increased evidently. CONCLUSION: HUVECs were induced to develop tubular structure in vitro cultured with Matrigel. GM-CSF promotes human vascular endothelial cells to form vessel-like structure in vitro in a dose-dependent and time-dependent manner. VEGF also in vitro promotes human vascular endothelial cells to form new vessel-like structure.     

VEGF induces HUVECs to produce extracellular H2O2 and its proliferation role

AIM:To study the effect of VEGF on extracellular H₂O₂ production in HUVECs and the role of H₂O₂ in the VEGF-induced proliferation. METHODS:HUVECs was stimulated with 500 μg/L VEGF. Products of extracellular H₂O₂ was detected by H₂DCFDA staining. MTT method was used to value the influences of 3×10⁶ U/L catalase and 5-20 mmol/L H₂O₂ to VEGF function. RESULTS:After treatment for 15 min with VEGF, HUVECs appeared fluorescence, and continued to become stronger, peaked at 45 min then decreased. HUVECs, which was treated simultaneity with VEGF and 3×10⁶ U/L catalase, only appeared very faint fluorescence. The proliferation of HUVECs by VEGF was restrained when treated with 3×10⁶ U/L catalase. The extrinsic H₂O₂ at concentration of 5-10 mmol/L promoted the proliferation of HUVECs but inhibited the proliferation effect of VEGF on HUVECs (P<0.01). CONCLUSION:These findings indicate that VEGF induces HUVECs to produce extracellular H₂O₂ and plays role in proliferation, but extrinsic H₂O₂ restrains VEGF function.     

Increased expression and possible roles of nicotinamide N-methyltransferase in pancreatic islets of STZ-induced diabetic monkeys

AIM: To verify and localize the expression of nicotinamide N-methyltransferase (NNMT) in pancreas of streptozotocin(STZ)-induced diabetic monkeys and understand its important role in β-cell destruction in the pathogenesis of diabetes. METHODS: Through an olig-microarray gene chip, NNMT was identified as the most obviously up-regulated genes in pancreas of STZ-induced diabetic monkeys versus controls. Semiquantitative RT-PCR and Western blotting were performed to verify the differential expression at mRNA and protein level respectively. Then the cellular localization of NNMT expression within pancreas was identified by immunohistochemical(IHC) staining.RESULTS: An obvious high expression of NNMT at both mRNA and protein levels was shown in pancreas of STZ-induced diabetic monkeys compared to that of controls. Further localization of the protein by IHC staining in pancreas specimens showed that its altered expression was restricted to central islets, most of which were β cells.CONCLUSION: Expression of NNMT is increased in islets of STZ- induced diabetic monkeys, which infers that NNMT might participate in the process of β cell damage in diabetes probably through the mechanism of energy metabolism disturbance.     

Effect of Ganoderma lucidum spores powder on the expression of IGF-1, NF-κB and apoptosis of nerve cells in the brain from epileptic rat

AIM: To investigate the effect of Ganoderma lucidum spores powder on the expression of insulin-like growth factor-1 (IGF-1), nuclear factor-κB (NF-κB) and apoptosis of nerve cells in rats with epilepsy established by pentetrazole. METHODS: The sub-eclampsia dosage of pentetrazole (PTZ) was used to make epilepsy model. Ganoderma lucidum spores powder group was given from stomach. The enduring time and latent period were recorded. The immune reactivity of IGF-1, NF-κB/P65 and apoptosis of nerve cells were measured with immunohistochemical staining and TUNEL method. RESULTS: In high power sight (×400), there were much more apoptosis cells in hippocampus and brain cortex of model group (18.80±2.13, 16.87±2.00) than those in control group (0.97±0.52, 0.58±0.25). The expressions of IGF-1, NF-κB in model group were higher than those in control group. Compared with model group, the latent period of Ganoderma lucidum spores powder group at the 17th, 21th, 25th days were longer (P<0.05, P<0.05, P<0.01, respectively).There were less apoptosis cells in hippocampus and brain cortex of Ganoderma lucidum spores powder group (12.30±2.46, 10.48±1.33) than those in model group. The expression of NF-κB/P65 in Ganoderma lucidum spores powder group was lower than that in model group, but the immune reactivity of IGF-1 increased more distinctly in Ganoderma lucidum spores powder group than that in model group. CONCLUSION: IGF-1, NF-κB and apoptosis of nerve cells may have play a role in occurrence and development of PTZ-induced epilepsy. Ganoderma lucidum spores powder can suppress expression of NF-κB strongly, and facilitate the immune reactivity of IGF-1, which may be one of the mechanisms by which Ganoderma lucidum spores powder restrains the apoptosis of nerve cells caused by epilepsy to prevent the damages of nerve cells.