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.
Cell Survival ; China ; Flowers ; physiology ; Fruit ; growth & development ; Orchidaceae ; growth & development ; physiology ; Pollen ; growth & development ; Reproduction ; Temperature

To examine whether or not the regulatory sequence of chicken ovalbumin gene can drive transgene expression specifically in hen oviduct, the authors constructed an oviduct-specific expression vector (pOV), containing 3.0 kilobases (kb) of the 5'-flanking sequence and 3.0 kb of the 3'-flanking sequence of the chicken ovalbumin gene. Jellyfish green fluorescence protein (EGFP) reporter gene and bacterial LacZ reporter gene were respectively inserted into the downstream of the 5'-regulatory region.The recombinants were named as pOVEGFP and pOVLacZ. Two transfer systems, in vitro and in vivo, were used to verify the function of the vector. In vitro, the plasmid DNA pOVEGFP and pEGFP-N1 were transfected respectively by the polyethyleneimine procedure into the primary chicken oviduct epithelium (PCOE) and fibroblasts cells isolated from laying hens. In vivo, the recombinant vector pOVLacZ was injected into egg-laying hens via wing vein and the tissues were collected for RT-PCR analysis.The results showed that expression of pEGFP-N1 was achieved at low level in oviduct epithelial cells and at high level in fibroblasts, but that the recombinant vector was not expressed in both cells. RT-PCR analysis showed that the LacZ gene was transcribed in the oviduct, but not in the heart, liver, kidney and spleen of the injected hens. Accordingly, the ?-galactosidase activity was only detected in the oviduct magnum (116.7 mU/ml) and eggs (16.47 mU/ml). These results indicated that the cloned regulation regions of chicken ovalbumin gene could drive exogenous gene expression specifically in the oviducts of hens. In vivo gene injection via wing vein may serve as a rapid production system of recombinant proteins in chicken eggs. In addition, the cultured primary oviduct cells from laying hens were not efficient temporary expression systems for analyzing the function of regulating elements of ovalbumin gene.

Collagen Type I ; analysis ; Collagen Type III ; analysis ; Fluorescent Antibody Technique, Indirect ; Humans ; Liver ; chemistry ; Microscopy, Confocal

To examine whether or not the regulatory sequence of chicken ovalbumin gene can drive transgene expression specifically in hen oviduct, the authors constructed an oviduct-specific expression vector (pOV), containing 3.0 kilobases (kb) of the 5'-flanking sequence and 3.0 kb of the 3'-flanking sequence of the chicken ovalbumin gene. Jellyfish green fluorescence protein (EGFP) reporter gene and bacterial LacZ reporter gene were respectively inserted into the downstream of the 5'-regulatory region. The recombinants were named as pOVEGFP and pOVLacZ. Two transfer systems, in vitro and in vivo, were used to verify the function of the vector. In vitro, the plasmid DNA pOVEGFP and pEGFP-N1 were transfected respectively by the polyethyleneimine procedure into the primary chicken oviduct epithelium (PCOE) and fibroblasts cells isolated from laying hens. In vivo, the recombinant vector pOVLacZ was injected into egg-laying hens via wing vein and the tissues were collected for RT-PCR analysis. The results showed that expression of pEGFP-N1 was achieved at low level in oviduct epithelial cells and at high level in fibroblasts, but that the recombinant vector was not expressed in both cells. RT-PCR analysis showed that the LacZ gene was transcribed in the oviduct, but not in the heart, liver, kidney and spleen of the injected hens. Accordingly, the beta-galactosidase activity was only detected in the oviduct magnum (116.7 mU/ml) and eggs (16.47 mU/ml). These results indicated that the cloned regulation regions of chicken ovalbumin gene could drive exogenous gene expression specifically in the oviducts of hens. In vivo gene injection via wing vein may serve as a rapid production system of recombinant proteins in chicken eggs. In addition, the cultured primary oviduct cells from laying hens were not efficient temporary expression systems for analyzing the function of regulating elements of ovalbumin gene.
Animals ; Cells, Cultured ; Chickens ; Cloning, Molecular ; methods ; Fallopian Tubes ; metabolism ; Female ; Organ Specificity ; Ovalbumin ; biosynthesis ; genetics ; Protein Engineering ; methods ; Recombinant Proteins ; biosynthesis ; Transfection ; methods ; Women

Background Indirect traumatic optic neuropathy (TON) is an acute injury of the optic nerve associated with severe visual dysfunction,which may be a result of secondary mechanical injury and vascular disorder of the optic nerve due to trauma.We analyzed the natural course of axonal loss and blood flow disturbances in patients with indirect TON to find a possible therapeutic window.Methods A cohort of 54 patients with indirect TON recruited between October 2008 and October 2010 at Beijing Tongren Hospital was retrospectively analyzed.The patients were divided into no light perception group (NLP) and better than NLP (btNLP) group.Specifically,the thickness of the retinal nerve fiber layer (RNFL) measured by spectral domain optical coherence tomography (SD-OCT),and hemodynamic parameters of the ophthalmic artery (OA),central retinal artery (CRA) and posterior ciliary artery (PCA) were determined.Results Two weeks after injury,there was a statistically significant decrease in the thickness of RNFL in the btNLP group as compared with the fellow control eyes (P ＜0.05).In contrast,in the NLP group,RNFL thickness slightly increased for 2 weeks following injury,then overtly reduced after 4 weeks (P ＜0.05).Peak systolic velocity (PSV) of CRA was significantly decreased 4 weeks after injury (P ＜0.05) in both the NLP group and btNLP group (P ＜0.05).The thickness of RNFL in the NLP group was negatively correlated with PSV of CRA after 1 week of injury (P ＜0.05,r=-0.962).Conclusions SD-OCT is a useful supplement in detecting the axonal loss in TON.The dynamic change of the thickness of RNFL appears to correlate with the hemodynamic disturbances in the natural course of TON.The first 2 weeks following an injury is critical and should be considered as the therapeutic window for TON patients.

Objective To evaluate the techniques and clinical effect of minimally invasive transfrontal puncture drainage using soft drainage tubes in the treatment of hypertensive basal ganglia hemorrhage. Methods Seventy-six patients with hypertensive basal ganglia hematomas received minimally invasive puncture drainage of the hematomas using soft drainage tubes. The puncture site was located in the frontal region on the hemorrhage side at 7-9 cm from the glabella and 3.5-4.5 cm adjacent to the midline. The puncture direction and depth was estimated according to the head CT data. Results The indwelling time of the drainage tubes ranged from 36 to 120 h (mean 70±5.6 h) after the operation. Three patients (3.95%) had recurrent hemorrhage after the operation, but intracranial infection or pneumocephalus occurred in none of the cases. Of tbe 76 patients, 11 had basic clinical cures, 35 showed marked improvements, and 16 showed improvements, with a total effective rate of 81.6%. Four patients failed to show improvements or had exacerbations, and 10 died after the operation, with a mortality rate of 13.2%. Conclusion Minimally invasive transfrontal puncture drainage using soft drainage tubes is effective for treatment of hypertensive basal ganglia hematomas. With simple surgical procedures, this approach can effectively reduce the occurrence of complications and improve the prognosis of the patients.

The catalytic generation of H2 in living cells provides a method for antioxidant therapy.In this study,an[FeFe]-hydrogenase mimic[Ru+Fe2S2@F127(80)]was synthesized by self-assembling polymeric plur-onic F-127,catalytic[Fe2S2]sites,and photosensitizer Ru(bpy)32+.Under blue light irradiation,hydrated protons were photochemically reduced to H2,which increased the local pH in living cells(HeLa cells).The generated H2 was subsequently used as an antioxidant to decrease reactive oxygen species(ROS)levels in living cells(HEK 293T,HepG2,MCF-7,and HeLa cells).Our findings revealed that the proliferation of HEK 293T cells increased by a factor of about six times,relative to that of other cells(HepG2,MCF-7,and HeLa cells).Intracellular ROS and pH levels were then monitored using fluorescent cell imaging.Our study showed that cell imaging can be used to evaluate the ability of Ru+Fe2S2@F127 to eliminate oxidative stress and prevent ROS-related diseases.

The longitudinal study sought to examine the dynamic development of cognitive skills for reading among elementary-level students in Mainland China.Two groups of students in first (n=164,mean age=6.65 years at first test) and second grade (n=202,mean age=7.73 years at first test) were followed on orthographic awareness,morphological awareness and rapid automatized naming (RAN) for two years.The children exhibited significant improvement in orthographic awareness,morphological awareness and RAN from grades 1 to 4.More importantly,to the orthographic and morphological awareness,while the children took a leap from grade 1 to 2 and grade 3 to 4,the progress developed at relatively slow rates from grade 2 to 3.In order to assure children's development of orthographic and morphological awareness,evidence-based orthographically and morphologically enhanced instruction is needed for Chinese children in the early elementary years,especially for those at the stage from grade 2 to 3.

Objective To investigate the effect of methylprednisolone sodium succinate on oxygen indexes (PaO2/FiO2) and serum biochemical indexes in patients with respiratory distress syndrome.Methods Seventy-two patients with respiratory distress syndrome were randomly divided into treatment group and control group,36 cases in each group.Control group was given routine treatment.Treatment group was given methylprednisolone sodium succinate 120 mg · d-1,by intravenous drip on the basis of control group.All the patients were treated continuously for two weeks.The PaO2/FiO2 and acute physiology and chronic health evaluation Ⅱ (APACHE Ⅱ) scores before and after treatment were observed.The levels of nitric oxide (NO),interleukin-6 (IL-6),tumor necrosis factor-α (TNF-α),C-reactive protein (CRP) and adverse drug reactions in two groups were observed.Results After treatment,the PaO2/FiO2 levels in treatment group and control were (287.62 ±24.57),(213.21 ±25.67)mmHg,the APACHE Ⅱ scores were 11.23 ± 1.66,15.59 ± 2.21,all with significant difference (all P ＜ 0.05).The levels of NO were (58.88±32.76) and (74.28 ±32.94) μ mol· L-1,the levels of IL-6 were (167.94 ±25.29) and (213.11±21.50) pg· mL-1,the levels of TNF-α were (22.35 ±8.93) and (36.30±9.88) pg·mL-1,the levels of CRP were (6.43 ± 3.02) and (9.25 ± 2.94) mg · L-1,all with significant differences (all P ＜ 0.05).There were 2 cases of short-term elevated blood pressure in treatment group,and there was no adverse drug reaction in control group,without significant difference (P ＞ O.05).Conclusion Methylprednisolone sodium succinate in the treatment of respiratory distress syndrome can significantly improve the PaO2/FiO2,reduce the APACHE Ⅱ score and serum biochemical indicators,and can improve the clinical effect with high safety.

OBJECTIVETo investigate the differentially expressed genes of proliferating and involuting hemangiomas by cDNA microarray analysis of gene-expression profiles in an effort to identify the key disease-related genes.

METHODSSamples were processed from total RNA and purified to mRNA, which was reverse-transcripted and hybridized onto Biodoor Genechip expression microarrays. Analyses were performed to determine the consensus pattern of gene expression in the proliferating and involuting stages of the same hemangioma and the changes in the expression level.

RESULTSIn proliferating hemangioma, 79 genes were overexpressed, and 115 genes were underexpressed in comparison with the involuting hemangioma. Some cytokines and growth factors such as neurotensin, Nov, CYR6, keratinocyte growth factor, interleukin-10 were overexpressed in proliferative hemangioma. In involuting hemangioma, apoptotic factors such as bcl-2 binding component, cytochrome C were overexpressed. The overexpression of Nov, CYR6, c-myc implied that angiogenesis and oncogenes might participate in the pathogenesis of hemangiomas. Mitochondria activated apoptotic passage (cytokines, bcl-2, cytochrome C) and Wnt/beta-catenin passage(Frizzled, beta-catenin, c-myc) were involved.

CONCLUSIONThe development of hemangiomas may be the results of imbalance of cell proliferation and apoptosis.

Apoptosis ; Cell Division ; Female ; Gene Expression Profiling ; Hemangioma ; genetics ; pathology ; Humans ; Male ; Oligonucleotide Array Sequence Analysis