Antitumor effects of different Ganoderma lucidum spore powder in cell- and zebrafish-based bioassays / 中西医结合学报

OBJECTIVE@#Ganoderma lucidum spore (GLS) is gaining recognition as a medicinal part of G. lucidum and has been reported to possess various pharmacological properties, such as antitumor activity. In this work, wall-broken GLS powder (BGLSP) and wall-removed GLS powder (RGLSP), two kinds of GLS powder with different manufacturing techniques, were compared in terms of contents of active constituents and in vivo and in vitro antitumor effects.@*METHODS@#The ultraviolet and visible spectrophotometry method was used to determine the contents of polysaccharides and total triterpenoids in BGLSP and RGLSP. Seventeen individual triterpenoids were further quantified using ultra-high-performance liquid chromatography and quantitative analysis of multi-components by single marker. The antitumor effects of BGLSP and RGLSP were evaluated using in vitro cell viability assay against human gastric carcinoma SGC-7901, lung carcinoma A549 and lymphoma Ramos and further validated by in vivo zebrafish xenograft models with transplanted SGC-7901, A549 and Ramos.@*RESULTS@#The results showed that the contents of polysaccharides, total triterpenoids and individual triterpenoids of RGLSP were significantly higher than those of BGLSP. Although both BGLSP and RGLSP inhibited the three tumor cell lines in vitro in a dose-dependent manner, the inhibitory effects of RGLSP were much better than those of BGLSP. In the in vivo zebrafish assay, RGLSP exhibited more potent inhibitory activities against tumors transplanted into the zebrafish compared with BGLSP, and the inhibition rates of RGLSP reached approximately 78%, 31% and 83% on SGC-7901, A549 and Ramos, respectively.@*CONCLUSION@#The results indicated that the antitumor effects of GLS were positively correlated with the contents of the polysaccharides and triterpenoids and demonstrated that the wall-removing manufacturing technique could significantly improve the levels of active constituents, and thereby enhance the antitumor activity.

Research advances in the relationship between iron deficiency and neurodevelopment in preterm infants / 中国当代儿科杂志

Iron deficiency (ID) is the most common micronutrient deficiency in children. Due to insufficient iron storage at birth and rapid catch-up growth after birth, preterm infants tend to have a high incidence rate of ID. During the critical period of brain development, ID alters iron-dependent neurometabolism, neurochemistry, neuroanatomy, and gene/protein profiles. This affects the central nervous system and causes the change in neurocognitive and behavioral development. Iron supplementation in infancy cannot reverse neurodevelopmental impairment caused by perinatal ID. The influence of ID on neurodevelopment is time- and region-specific, and in the high-risk population, early diagnosis and optimal iron treatment may help with the recovery of brain function and improve quality of life and long-term prognosis in preterm infants.

Role of vasodilator stimulated phosphoprotein in resistin-induced proliferation and migration of human coronary endothelial cells / 医学研究生学报

Objective Resistin,also known as an adipose tissue-specific secretory factor,is involved in the development and progression of artherosclerosis by inducing dysfunction of endothelial cells,but its action mechanism has been rarely studied.The purpose of this study was to investigate the effect of resistin on the function of human coronary artery endothelial cells (HCAECs) and the underlying mechanisms.Methods We constructed the lentiviral vasodilator-stimulated phosphoprotein (VASP) shRNA interfering sequence (the LV-siVASP group) and a negative control vector (the LV-sicntr group),transfected HCAECs with VASP siRNA or control siRNA,and determined the VASP mRNA and protein expressions by quantitative RT-PCR and Western blot,respectively.We treated the HCAECs with resistin at the concentrations of 0,10,50,100,and 200 ng/mL followed by-detection of its effects on the proliferation and migration of the cells by MTT and Transwell chamber assay,respectively.We made comparisons between the LV-siVASP and LV-sicntr groups in the proliferation and migration of the HCAECs treated with 100 ng/mL resistin,the expression of vascular endothelial growth factor receptor-2 (VEGFR2) by immunofluorescent staining,and the activity of RhoA by pull-down assays.Results Compared with the LV-sicntr group,the LV-siVASP group showed significantly inhibited expressions of RNA (100％ vs [68.1±0.8]％,P＜0.05) and the VASP protein (100％ vs [59.3± 1.7] ％,P＜0.05) Treatment with resistin at 50-200 ng/mL markedly increased the proliferation of the HCAECs at 48 hours (P＜0.05) and induced a dose-dependent promotion of their migration at 24 hours (P＜0.05).No significant difference was observed in the expression VEGFR2 between the two groups,while the activity if RhoA was remarkably reduced in the LV-siVASP group ([41.3±3.1] ％) as compared with the LV-sicntr group (P＜0.05).Conclusion Resistin promotes the proliferation and migration of HCAECs,and the influence of VASP ablation on the action of resistin is associated with the decreased activity of RhoA.

Pollen viability, stigma receptivity and fruiting characteristics of botanical origin of Jinxianlian / 中国中药杂志

The viability and life span of pollen were evaluated by TTC (2,3,5-triphenyl tetrazlium chloride) and the peroxidase solution, the stigma receptivity were estimated by benzidine-H2O2 method and the fruiting characteristics were investigated. The results showed that (1) Anoectochilus roxburghii and A. formosanus appeared the same up-and-down trend of the pollen viability, increased and then decreased. The storage temperature and storage time had significant impact on the pollen viability. With the extension of storage time, the pollen activity decreased. 4 degrees C refrigerator storage may be extended the pollen vitality. (2) The stigma had receptivity in 1st day and reached the highest level in the 4th day after blooming. A. roxburghii lost receptivity in the 8th day while A. formosanus lost receptivity in the 10th day after blooming. (3) The different pollination had significant impact on seed setting rate. The seed setting rate of artificial cross-pollination was higher than that of the artificial self-pollination. Collecting pollen in the 3rd day and carrying out artificial cross-pollination in the 4th day after blooming can significantly improve seed setting rate. The results provided technical assurance for A. roxburghii and A. formosanus breeding of new varieties and seed breeding.

Influence of seedling grade on plant growth, yield and quality of Anoectochilus roxburghii / 中国中药杂志

The morphological index of the seedlings including the plants height, the ground diameter, the leaf amounts, the fresh weight of the whole plant and the ratio of height to diameter was measured and the principal components were analyzed so as to determine the grading index, and stepwise cluster analysis was applied for clustering analysis. Pot experiments were used to measure the indicators of plant growth and development, the yield and the quality. The results showed that the height and ground diameter were determined as the quality indicators of the seedlings grading and the standard quality grading of seedlings of Anoectochilus roxburghii was initially set up, different seeding plants influenced the plants growth and the yield. The ground diameter of the class I was larger than that of the class II and III, so as the yield. The seedling grading had no obvious effect on the internal quality of medicinal materials. The results of the study provide the basis for standard cultivation of A. roxburghii.

Effects of different transplanting conditions on survival rate and growing status of Anoictochilus roxburghii plantlets / 中国中药杂志

The growing status of Anoectochilus roxburghii seedling was observed and the survival rate of seedlings, height, stem diameter and plant fresh weight under the conditions of different transplanting substrate compositions, planting density, shading rate were measured. The results showed that the effects of different transplanting substrates, planting densities, shading rates and nutrient solutions on the growing status of A. roxburghii plantlets varied greatly. A. roxburghii plantlets demonstrated a high survival rate and better growing status under the Following conditions: the ratio of peat and river sand as 2: 1, the planting density as 3 cm x 3 cm, the shading rate as 70%, and the nutrient solution as 1/4MS. The findings of the study provide a solid technical solution for the artificial cultivation of A. roxburghii plantlets.

Biological characteristics of Fusarium oxysporum and inhibitory effects of five fungicides / 中国中药杂志

The mycelium growth and sporulation in different temperature, pH value and light conditions were detected by using the crossing and haemocytometer method. The toxicity of five fungicides to Fusarium oxysporum was tested by mycelia growth method, so as to provide the reference for prevention and control against F. oxysporum. The optimum temperature and pH value of F. oxysporum to grow and spore were 28 degrees C and 6-7. Alternating light and darkness promoted growth and sporulation of bacterial colony. As for five fungicides, the EC50 of tebuconazole was 10.02 mg x L(-1), 92.50 times as much as carbendazim. The EC50 of myclobutanil and Fuxing was 91.23, 96.68 mg x L(-1), respectively. Tebuconazole showed the most tremendous inhibitory effect and control efficiency on F. oxysporum.

Studies on methods of in vitro evaluation of soft rot-resistance in jinxianlian / 中国中药杂志

Compared with living spray method, it focused on the investigation of different inoculation methods, various inoculation concentration and the influence of different seeding age on soft rot-resistance in Jinxianlian. The results showed that (1) Inoculated with dropping connection, the difference of disease index between A. roxburghii and A. formosanus was grate, so that the disease-resistance could be obviously distinguished. (2) When the inoculation concentration was 1.0 x 10(7) cfu x mL(-1), the difference of disease index was relatively obvious and the disease-resistance could be differentiated well. (3) At the moment of 4-month seeding inoculation, a certain difference of the disease index between A. roxburghii and A. formosanus was existed, so, relatively, it could accurately reflect the resistance difference between various species. With the inoculation of dropping connection, A. roxburghii and A. formosanus of 4-month seeding age was put in the bacteria suspension of inoculation concentration of 1.0 x 10(7) cfu x mL(-1). The identification was taken up after 5 days in the incubator under the condition of 14 h daylight and 28 degrees C. The identification result was conformed with that of the living spray method. To investigate the identification method of in vitro evaluation of soft rot-resistance of Jinxianlian so as to provide the foundation for germplasm utilization and excellent cultivars breeding.

Residue decline dynamics and safety utilization of carbendazim in cultivation of Anoectochilus roxburghii / 中国中药杂志

The paper aimed to study the residue decline dynamic and standards for safety utilization of carbendazim in roots, stems, leaves of Anoectochilus roxburghii and in growth media. Samples extracted with methanol were purified by liquid-liquid extraction and analysed by HPLC. The results showed that average rate of recovery was 82.9% - 95.7% and RSD were 2.0% - 6.3% with add of carbendazim in respectively diverse concentration, which meets inspection requirement of pesticide residue. Two kinds of dosages of carbendazim were treated, varying from recommended dosage (1.0 kg x hm(-2)) to 1.5 times recommended dosage (1.5 kg x hm(-2)). Results of two years test showed that the half-life period of carbendazim were 7.01 - 8.51 d in the growth media of A. roxburghii, 3.58 - 4.27 d in stems and 3.50 - 3.91 d in leaves, 4.93 - 5.71 d in roots. Providing max recommended residue of carbendazim in the cultivation of A. roxburghii is 0.5 mg x kg(-1), sprayed 4 times a year with the dosage of 1.0 kg x hm(-2), 28 days is proposed for the safety interval of the last pesticide application's and harvest's date.

Multiple analysis of relationship between morphologic traits and yield formation of Anoectochilus roxburghii / 中国中药杂志

Nine morphologic traits, plant height, ground diameter, leaf length, leaf width, leaf area of plant, leaf fresh weight, blades, length/width ratio, plant fresh weight of Anoectochilus roxburghii from 13 different areas were determined for correlation analysis, path analysis and principal components analysis. Different source of morphological trait variation coefficient of A. roxburghii was 2.96% -12.59%, plant fresh weight was significant positively correlated with ground diameter, plant height and leaf number, and positively correlated with leaf fresh weight. Path analysis showed that plant height had the largest positive direct effect on plant fresh weight, the leaf fresh weight and blades number had indirect effects on the plant fresh weight. Through principal component analysis, morphological traits of A. roxburghii can be divided into "Determinants of high-yielding morphology" and "Determinants of leaf production". In the actual process of production and breeding of A. roxburghii, we should pay attention to plant height, leaf fresh weight, blades numbers and other traits.

Breeding of Dendrobium officinale var. 'Xianhu 2' and its characteristics / 中国药学杂志

OBJECTIVE: To select and breed elite variety of Dendrobium officinale, thus to conserve the wild germplasms and promote the healthy developing of D. officinale industry. METHODS: System breeding method was employed for the breeding of new variety developed from wild germplasm oi Dendrobium officinale. Yield of fresh stems and leaves, quality characters including dried stem rate and raw polysaccharide content etc. were tested. ISSR molecular marker technique was used to distinguish genetic difference between new variety and other D. officinale materials. RESULTS: New elite variety 'Xianhu 2' was bred successfully with high polysaccharide content, fine adaptability, and high yield. 'Xianhu 2' had significantly higher yields of fresh stems-leaves and stems than D. officinale var. 'Yunnan Ruanjiao' and D. officinale var. 'Xianhu 1' and significantly higher dried stem ratio and raw polysaccharide content than 'Yunnan Ruanjiao' at 12, 24, and 36 m, respectively. ISSR analysis indicated that there were distinct differences in DNA level between 'Xianhu 2' and other D. officinale varieties and lines. D. officinale var. 'Xianhu 2' got its certificate for commerce releasing by Zhejiang Province in December 2011. CONCLUSION: 'Xianhu 2' is a new elite variety with good synthetic agronomic traits. Its breeding is beneficial for the cultivation of D. officinale.

Controlled clinical trials on different surgical methods for the treatment of acromioclavicular dislocation / 中国骨伤

<p><b>OBJECTIVE</b>To compare clinical effects of clavicular hook plate fixation, coracoid transplantation, and clavicular hook plate fixation combined with modified dynamic muscle transfer for the treatment of the complete acromioclavicular dislocation.</p><p><b>METHODS</b>From January 2006 to November 2009, 65 patients with sustained complete acrominoclavicular dislocation were treated with clavicular hook plate fixation, coracoid transplantation,and clavicular hook plate fixation combined with modified dynamic muscle transfer. All the patients were divided into three groups: 22 patients in group A were treated with clavicular hook plate fixation, including 17 males and 5 females, with an average age of (31.0 +/- 10.0) years; 21 patient in group B were treated with coracoid transplantation, including 16 males and 5 females,with an average age of (33.0 +/- 6.4) years; 22 patients in group C were treated with clavicular hook plate fixation combined with modified dynamic muscle transfer,including 18 males and 4 females, with an average age of (30.0 +/- 5.3) years. Postoperative functional recovery was evaluated by Karlsson criteria.</p><p><b>RESULTS</b>All the patients were followed up, and the duration ranged from half to three years (averaged 1.5 years). In group A, 8 patients got half re-dislocation, 2 patients got complete re-dislocation and arthritis of acromioclavicular joint after internal fixations removal, 1 patient had clavicular hook plate broken after operation. In group B, 7 patients got half re-dislocation, 1 patient got complete re-dislocation,and 5 patients had arthritis of acromioclavicular joint with acute pain and limited shoulder function after internal fixations removal. In group C,2 patients got half re-dislocation, no complete re-dislocation and arthritis of acromioclavicular joint occurred after internal fixations removal. According to Karlsson evaluation, in group A, 12 patients obtained an excellent result, 8 good and 2 poor; in group B, the data were 9, 7 and 5 respectively; in group C, they were 20, 2 and 0 respectively. There were remarkable differences of therapeutic effects between the clavicular hook plate fixation combined with modified dynamic muscle transfer and that with either of the former two treatment methods (P < 0.05).</p><p><b>CONCLUSION</b>Clavicular hook plate combined with modified dynamic muscle transfer is a reliable and good treatment for the complete acrominoclavicular dislocation, with advantages such as easy to handle,stable fixation and early exercise.</p>

Effects of recombinant adenovirus-mediated triple mutant of hypoxia inducible factor-1alpha in modulating angiogenesis / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the effect of recombinant adenovirus-mediated triple mutant of hypoxia inducible factor-1alpha (Ad-HIF-1alpha-564/402/803) in modulating angiogenesis in vitro.</p><p><b>METHODS</b>The recombinant adenoviruses Ad-lacZ, Ad-Null, Ad-HIF-1alpha-nature, and Ad-HIF-1alpha-564/402/803 were amplified in HEK293A cells and purified by ultracentrifugation in CsCl step gradient solutions, and the adenoviral titer was determined by end-point dilution assay. The recombinant adenovirus was confirmed by PCR and DNA sequence analysis, and the infection efficiency was observed by X-gal staining. Human microvascular endothelial cells (HMVECs) were infected with Ad- HIF-1alpha-564/402/803, Ad- HIF-1alpha-nature, or Ad-Null to compare the number of capillary-like tube structures in vitro. The effect of Ad- HIF-1alpha-564/402/803, Ad-HIF-1alpha-nature, and Ad-Null on angiogenesis was evaluated using a chick embryo chorioallantoic membrane (CAM) model.</p><p><b>RESULTS</b>PCR and gene sequencing suggested the correct construction of the recombinant adenovirus HIF-1alpha, and the adenoviral titer reached 1011-1012 PFU/ml. Infection of the hMVECs with Ad-HIF-1alpha-564/402/803 at the optimal multiplicity of infection of 100 pfu/cell resulted in a significantly greater number of capillary-like tube structures than infection by Ad-HIF-1alpha-nature and Ad-Null (P=0.000). Ad-HIF-1alpha-564/402/803 group showed significantly higher microvessel density than Ad-HIF-1alpha-nature, Ad-Null, and PBS groups, with also higher angiogenesis area to CAM area ratio (P=0.01, 0.000, and 0.000, respectively).</p><p><b>CONCLUSION</b>The triple mutant Ad-HIF-1alpha-564/402/803 can obviously promote the formation of capillary-like tube structures in vitro and modulate angiogenesis in the CAM model, suggesting the capacity of Ad-HIF-1alpha-564/402/803 in promoting angiogenesis under normoxic condition.</p>

Effect of different doses of recombinant adenovirus mediated hypoxia-inducible factor-1alpha on angiogenesis in ischemic lower hind limb of rabbits / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the effect of recombinant adenovirus-mediated hypoxia-inducible factor-1alpha (Ad-HIF-1alpha) at different doses on angiogenesis in a rabbit model of hind limb ischemia.</p><p><b>METHODS</b>Left hind limb ischemia was induced in 45 Zealand white rabbits by ligation of the left femoral artery. The rabbits were randomly divided into 5 groups (n=9) to receive intramuscular injections of 0.5 ml saline, 2x10(10) PFU empty vector (Ad-null), or different doses of Ad-HIF-1alpha (2x10(9), 2x10(10) or 2x10(11) PFU) immediately after the operation. On the 7th day after the operation, real-time PCR was used to detect the expression of HIF-1alpha mRNA in the skeletal muscles. Immediately and on the 14th and 28th days after the operation, contrast enhanced ultrasound (CEU) was used to observe the blood perfusion of the hind limb. On the 28th day postoperatively, immunohistochemistry for CD31 was performed to evaluate the microvascular density (MVD).</p><p><b>RESULTS</b>Real-time PCR showed that Ad-HIF-1alpha significantly increased the expression of HIF-1alpha mRNA (P<0.01) in a dose-dependent manner as compared with that in the saline and Ad-null groups (P<0.01). CEU revealed greater blood perfusion in the hind limb of rabbits in association with increased dose of Ad-HIF-1alpha (P<0.05 or P<0.01); similar changes in the MVD was observed following Ad-HIF-1alpha injections as shown by immunohistochemistry (P<0.05 or P<0.01). No significant differences were found either in the blood perfusion or MVD between saline and Ad-null groups (P>0.05).</p><p><b>CONCLUSION</b>Ad-HIF-1alpha can dose-dependently promote the angiogenesis in the ischemic limb of rabbits.</p>