Improving precision in the assessment of round cell numbers in human semen

The aim of this study was to increase the precision of assessment of the number of round cells observed in the per- oxidase test for detection of seminal leukocytes （granulocytes）. The dilution of semen was reduced and the volume of suspension examined was increased for semen samples containing between 0.6 and 6 million round cells per mL. A 1 ＋ 5 （1：6） dilution was compatible with measurable peroxidase activity and a sufficiently clear background for cell assessment. At this dilution, and with assessment of all 18 grids on both sides of the Neubauer-improved counting chamber, only three of the 10 samples （nominal cell concentrations of 1.9 ×10^-6-3.3×10^-6 mL-1） presented 400 round cells or more. As lower seminal dilutions were incompatible with easy detection of round cells or their peroxidase reaction product, it was not pos- sible to provide precise measurements （sampling error 5%） of the suggested lower reference limit of 1 ×10^-6 cells per mL. The results indicate that this poor precision of measuring 1×10^-6 round cells per mL could explain the discrepant reports on the acceptability of the cut-off values for leukocytospermia. Such reference limits need to be established with statistically sound methods.

Cell number explains the intraspecific spur-length variation in an Aquilegia species

Variations of nectar spur length allow pollinators to utilize resources in novel ways,leading to the different selective pressures on spurs and allowing taxa to diversify.However,the mechanisms underlying spur length variation remain unclear.Interspecific comparisons of spur length suggest that both cell division and anisotropic expansion could explain the changes of spur length,and that hormone-related genes contribute to the process of spur formation.In contrast,little is known about intraspecific spur length variation.In Aquilegia rockii,spur length varies strikingly,ranging from 1 mm to 18 mm.To examine the potential mechanisms underlying spur length variation in A.rockii,we observed cell morphology and analyzed RNA-seq of short-and long-spurred flowers.Scanning electron microscopy revealed that at two positions on spurs there were no differences in either cell density or cell anisotropy between short-and long-spurred flowers,suggesting that in A.rockii changes in cell number may explain variations in spur length.In addition,we screened transcriptomes of short-and long-spurred flowers for differentially expressed genes;this screen identified several genes linked to cell division(e.g.,F-box,CDKB2-2,and LST8),a finding which is consistent with our analysis of the cellular morphology of spurs.However,we did not find any highly expressed genes involved in the hormone pathway in long-spurred flowers.In contrast to previous hypotheses that anisotropic cell expansion leads to interspecific spur variation in Aquilegia,our results suggest that cell number changes and related genes are mainly responsible for spur length variations of A.rockii.Furthermore,the underlying mechanisms of similar floral traits in morphology may be quite different,enriching our understanding of the mechanisms of flower diversity in angiosperms.

Pollination dynamics, grain weight and grain cell number within the inflorescence and spikelet in oat and wheat

Oat (Avena sativa L.) and wheat (Triticum aestivum L.) vary in the structure of their inflores-cences and also in how pollination proceeds within the inflorescence. In both species the grain position in the spikelet determines grain weight potential. Primary grains in oat and proximal grains in wheat weigh more than secondary and distal grains. This variation in grain weight can potentially result from differences in post-pollination cell division in the grain. In this study pollination duration and dynamics were analyzed from head samples collected at two-day intervals, starting from the pollination of the most advanced floret. The number of grain cells was determined for individual grains throughout the inflorescence, starting from the pollination event. When mature, grain position in the spikelet and spike was noted and grain weight assessed. Pollination advance in oat proceeded from the uppermost primary floret towards the basal spikelets in ten to eleven days. Within the spikelet, the primary floret was pollinated on average one day earlier than the secondary floret. In wheat, pollination duration was four to five days, starting from the proximal florets in the mid-section of the inflorescence progressing towards the apical and basal spikelets. Proximal florets were pollinated one to two days earlier than distal florets. Maximum cell number in primary grains exceeded that of secondary grains in two oat cultivars. Similarly, primary grains were heavier than secondary grains. Cell number and single grain weight were correlated in terms of grain position in the spikelet (primary – secondary) and cultivar. Oat cultivar Belinda had a higher single grain weight than Fiia, which was also expressed as larger grain cell number. In wheat, proximal grains had higher maximum cell numbers and were also heavier than distal grains. This grain weight gradient was apparent throughout the inflorescence. Consequently, grain cell number is one of the possible regulators of grain-filling capacity in both cereal crops.

Hi-Tag:a simple and efficient method for identifying protein-mediated long-range chromatin interactions with low cell numbers

Protein-mediated chromatin interactions can be revealed by coupling proximity-based ligation with chromatin immunoprecipitation.However,these techniques require complex experimental procedures and millions of cells per experiment,which limits their widespread application in life science research.Here,we develop a novel method,Hi-Tag,that identifies high-resolution,long-range chromatin interactions through transposase tagmentation and chromatin proximity ligation(with a phosphorothioate-modified linker).Hi-Tag can be implemented using as few as 100,000 cells,involving simple experimental procedures that can be completed within 1.5 days.Meanwhile,Hi-Tag is capable of using its own data to identify the binding sites of specific proteins,based on which,it can acquire accurate interaction information.Our results suggest that Hi-Tag has great potential for advancing chromatin interaction studies,particularly in the context of limited cell availability.

Study on the Relationship Between the Ploidy Level of Microspore-Derived Plants and the Number of Chloroplast in Stomatal Guard Cells in Brassica oleracea

The relationship between the ploidy level of microspore-derived plants and chloroplast number in stomatal guard cells was studied in cabbage, broccoli, and Chinese kale. In the experiment, distribution statistics analysis and t-test were used to perform statistical analysis on chloroplast number of different ploidy level in those stomatal guard cells mentioned above, and morphology identifying and chromosome counting were used to test accuracy of counting chloroplast number in stomatal guard cells. The chloroplast average number in stomatal guard cells was very similar among the different leaf positions on the same plant and among significantly among the different ploidy the different locations in the same stoma in the same variety. All the leaf, while the chloroplast number varied distributions of the chloroplast number in different ploidy stoma were normal distribution fitted. A correlation has been established between ploidy and chloroplast number in the stomatal guard cells. In every single stoma of microspore-derived plants, the chloroplast number for a haploid should not be more than 10, diploids 11 to 15, and polyploids more than 15. The accuracy of this method for identification of different ploidy plants was 93.93%. Furthermore, the accuracy of this method was reliable and did not vary with the plants growth conditions. Therefore, the chromosome ploidy of plants derived from microspore culture in cabbage, broccoli, and Chinese kale can be identified by simply counting the chloroplast number in stomatal guard cells.

基于模糊n-cell数的非线性投入产出模型解的存在性

将混合单调以及耦合不动点的概念引入到模糊n-cell数空间中,给出此空间上的混合单调型不动点定理。考虑到模糊n-cell数具有表示n维不确定信息的特点,将非线性投入产出模型与模糊n-cell数相结合,建立模糊非线性投入产出模型并给出相应的平衡方程。最后利用本文中所给的不动点定理讨论此模型解存在的条件,验证模型的合理性。

Relationship between epidermal growth factor receptor gene mutation and copy number in Chinese patients with non-small cell lung cancer

Epidermal growth factor receptor(EGFR) gene mutation and copy number are useful predictive markers that guide the selection of non-small cell lung cancer(NSCLC) patients for EGFR-targeting therapy.This study aimed to investigate the correlation between EGFR gene mutation and copy number and clinicopathologic characteristics of Chinese patients with NSCLC.NSCLC specimens collected from 205 patients between November 2009 and January 2011 were selected to detect EGFR gene mutations with real-time polymerase chain reaction(RT-PCR) and to detect EGFR gene copy number with fluorescence in situ hybridization(FISH).EGFR mutations primarily occurred in females,non-smokers,and patients with adenocarinomas(all P < 0.001).Tissues from 128(62%) patients were FISH-positive for EGFR,including 37(18%) with gene amplification and 91(44%) with high polysomy.EGFR gene mutation was correlated with FISH-positive status(R = 0.340,P < 0.001).Multivariate analysis showed that not smoking(OR = 5.910,95% CI = 2.363-14.779,P < 0.001) and having adenocarcinoma(OR = 0.122,95% CI = 0.026-0.581,P = 0.008) were favorable factors for EGFR gene mutation.These results show a high frequency of EGFR FISH positivity in NSCLC tissues from Chinese patients and a significant relevance between EGFR gene mutations and FISH-positive status.Among the FISH-positive samples,EGFR gene mutation occurred more frequently in samples with gene amplification compared to those with high polysomy,suggesting that EGFR mutation and gene amplification should be used as clinical decision parameters to predict response to EGFR-targeting therapy.

Analysis of Syetem Reliability in Manufacturing Cell Based on Triangular Fuzzy Number

Due to lacking of test-data and field-data in reliability research during the design stage of manufacturing cell sys- tem.The degree of manufacturing cell system reliability research is increased.In order to deal with the deficient data and the un- certainty occurred from analysis and judgment,the paper discussed a method for studying reliability of manufacturing cell system through the analysis of fuzzy fault tree,which was based on triangular fuzzy number.At last,calculation case indicated that it would have great significance for ascertaining reliability index,maintenance and establishing keeping strategy towards manufac- turing cell system.

Prognostic value of MET, cyclin D1 and MET gene copy number in non-small cell lung cancer

The aim of this study was to analyze the correlation of the expression of MET and cyclin D1 and MET gene copy number in non-small cell lung cancer （NSCLC） tissues and patient clinicopathologic characteristics and sur- vival. Sixty-one NSCLC tissue specimens were included in the study. The expression of MET and cyclin D1 was evaluated by immunohistochemistry and MET gene copy number was assessed by quantitative real-time polymer- ase chain reaction （Q-PCR）. Positive expression of MET and cyclin D1 protein and increased MET gene copy number occurred in 59.0%, 59.0% and 18.0% of 61 NSCLC tissues, respectively. MET-positivity correlated with poor differentiation （P = 0.009）. Increased MET gene copy number was significantly associated with lymph node metastasis （P = 0.004） and advanced tumor stage （P = 0.048）, while the expression of cyclin D1 was not associ- ated with any clinicopathologic parameters. There was a significant correlation between the expression of MET and MET gene copy number （P = 0.002）. Additionally, the expression of cyclin D1 had a significant association with the expression of MET as well as MET gene copy number （P = 0.002 and P = 0.017, respectively）. MET- positivity and increased MET gene copy number were significantly associated with poor overall survival （P = 0.003 and P 〈 0.001, respectively） in univariate analysis. Multivariate Cox proportional hazard analysis confirmed that the expression of MET and MET gene copy number were prognostic indicators of NSCLC （P = 0.003 and P = 0.001, respectively）. The overexpression of MET and the increased MET gene copy number might be adverse prognostic factors for NSCLC patients. The activation of the MET/cyclin D1 signaling pathway may contribute to carcino- genesis and the development of NSCLC, and may represent a target for therapy.

The Effect of Different Surgical Methods on the Number of Circulating Tumor Cells in the Peripheral Blood of Patients with Renal Cell Carcinoma

Objective: The objective is to explore the effects of different surgical methods-retroperitoneal laparoscopic radical nephrectomy (RLRN) and open radical nephrectomy (ORN) on the number of circulating tumor cells (CTC) in the peripheral blood of patients with renal cancer. Methods: The clinical data of 63 patients in the Department of Urology, Affiliated Hospital of Chengde Medical College who underwent radical surgery for renal cancer were divided into CTC positive group (18 cases of open surgery and 16 cases of minimally invasive surgery) and CTC negative group (14 cases of open surgery), 15 cases of minimally invasive surgery), overall group (32 cases of open surgery, 31 cases of minimally invasive surgery). Observe the changes in the number of CTC 1 week before operation and 1 week after operation. Results: In the positive group, whether it was open surgery or minimally invasive surgery, the postoperative CTC level of patients was significantly reduced (P < 0.05). In the negative group, the CTC decreased significantly after open surgery (P > 0.05), and the CTC level decreased significantly after minimally invasive surgery (P < 0.01). In the overall group, both open and minimally invasive surgery CTC decreased significantly, and the difference was statistically significant (P < 0.05). Conclusion: The two different surgical methods can reduce the level of CTC, but compared with ORN, RLRN can significantly reduce the number of postoperative CTC. Patients in the CTC-negative group may be less suitable for open surgery. CTC levels have certain potential in the selection and guidance of treatment modes for patients with renal cell carcinoma (RCC).

基于OWA算子的模糊n-cell数排序方法

针对基于模糊n-cell数的多属性排序问题,提出了一种基于有序加权平均算子(OWA算子)的模糊n-cell数排序方法。该方法首先根据样本数据对评估对象的属性构造模糊n-cell数,其次根据均值将属性按照从大到小排列,然后选取合适的权重向量,应用OWA算子进行信息聚合得到综合模糊n-cell数,接着根据各分量均值得到排序结果。最后,将该方法运用到实例中,并与传统的均值方法进行了比较。结果表明该方法不仅灵活有效,可根据具体情况选择不同的OWA权重来消除部分不合理的情况,使结果更有说服力,还弥补了传统均值方法的不足。

The Difference of the Copy Number Variation and Loss of Heterozygosity of Human Lung Large Cell Cancer Cell Line with Different Metastatic Potential

Background and Objective It has been proven that copy number gain/or loss (copy number variation CNV) in uences gene expression and result in phenotypic variation by

Nerve growth factor promotes in vitro proliferation of neural stem cells from tree shrews

Neural stem cells promote neuronal regeneration and repair of brain tissue after injury,but have limited resources and proliferative ability in vivo.We hypothesized that nerve growth factor would promote in vitro proliferation of neural stem cells derived from the tree shrews,a primate-like mammal that has been proposed as an alternative to primates in biomedical translational research.We cultured neural stem cells from the hippocampus of tree shrews at embryonic day 38,and added nerve growth factor（100 μg/L） to the culture medium.Neural stem cells from the hippocampus of tree shrews cultured without nerve growth factor were used as controls.After 3 days,fluorescence microscopy after DAPI and nestin staining revealed that the number of neurospheres and DAPI/nestin-positive cells was markedly greater in the nerve growth factor-treated cells than in control cells.These findings demonstrate that nerve growth factor promotes the proliferation of neural stem cells derived from tree shrews.

直觉模糊n-cell数及其构造方法

基于一种特殊类型的n维模糊数——模糊n-cell数,定义了直觉模糊n-cell数,其可以更有效地处理高维不确定数据且便于应用.然后给出并证明了直觉模糊n-cell数的表现定理.最后,依据数据集的均值以及离散度设计了直觉模糊n-cell数的一种构造方法.

模糊n-cell数近似大于等于关系的性质

研究了模糊n-cell数近似大于等于关系、近似大于关系在水平收敛下的变化,与通常实数序列性质类似,近似大于等于关系在水平收敛下能保持不变,近似大于关系在水平收敛下无法保持。但是与通常实数列极限具有保号性不一样的是,水平收敛的模糊n-cell数序列关于近似大于关系的保号性不成立。

基于直觉模糊n-cell数的多属性决策方法及应用

针对权重已知的直觉模糊多属性决策问题,提出了一种基于直觉模糊n-cell数的决策方法。定义了直觉模糊n-cell数的偏序和两种基于均值和离散度的弱序。研究了所提出序的理论性质,并进行比较分析。最后,将相关排序方法应用于一类人才选拔的问题之中,在解决问题的同时验证了方法的有效性。

A comprehensive genomic characterization of esophageal squamous cell carcinoma: from prognostic analysis to in vivo assay

Background: Esophageal squamous cell carcinoma(ESCC) is a leading cause of cancer death worldwide and is char?acterized by numerous genetic mutations. TNM staging is not sufficient for predicting patient outcomes. Addition?ally, ESCC shows poor responsiveness to chemotherapy and radiation. Thus, there is an urgent need to find efficient therapy targets. Previous ESCC high?throughput genomic studies have lacked intensive survival analysis, particularly for copy number variation(CNV) and the genes involved.Main body: In the study "Genomic Characterization of Esophageal Squamous Cell Carcinoma Reveals Critical Genes Underlying Tumorigenesis and Poor Prognosis" recently published in the American Journal of Human Genetics, we comprehensively analyzed the effects of CNVs, mutations, and relative gene expression on patient outcomes. To validate our findings for our 67 sequencing samples, we collected a 321?patient retrospective cohort with detailed 5?year follow?up information and carried out univariate and multivariate survival analyses. In addition, the biological functions of the survival predictors in ESCC were investigated both in vitro and in vivo.Conclusions: We found the independent ESCC survival predictors and potential therapy targets. Nevertheless, the effects of numerous low?frequency mutations need to be explored using larger sample sequencing. Overall, con?structing multi?gene prognostic signatures will remain a great challenge in the future.

Enhancing endothelial progenitor cell for clinical use

Circulating endothelial progenitor cells(EPCs) have been demonstrated to correlate negatively with vascularendothelial dysfunction and cardiovascular risk factors. However, translation of basic research into the clinical practice has been limited by the lack of unambiguous and consistent definitions of EPCs and reduced EPC cell number and function in subjects requiring them for clinical use. This article critically reviews the definition of EPCs based on commonly used protocols, their value as a biomarker of cardiovascular risk factor in subjects with cardiovascular disease, and strategies to enhance EPCs for treatment of ischemic diseases.

Pyroelectric Bi_5-x(Bi₂S₃)₃₉I₁₂S: Fibonacci Superstructure, Synthesis Options and Solar Cell Potential

Previously, synthetic hexagonal bismuth sulfide iodide (polar space group P63, a = 15.629(3) ?, c = 4.018(1) ?, Z = 2) has been described by the rather unsatisfactory fractional formula Bi19/3IS9 [1] 08D0C9EA79F9BACE118C8200AA004BA90B02000000080000000E0000005F005200650066003400310037003600350038003400370039000000 -[3] 08D0C9EA79F9BACE118C8200AA004BA90B02000000080000000E0000005F005200650066003400310037003600350038003400370036000000 . A redetermination of the structure using old but reliable photographic intensity data indicated the presence of additional split positions and reduced atomic occupancies. From the observed pattern of this “averaged” structure a consistent model of a superstructure with lattice parameters of a' = √13·a = 56.35(1) ?, c' = c, and a formula Bi5-x(Bi2S3)39I12S emerged, with 2 formula units in a cell of likewise P63 space group. Structural modulation may be provoked by the space the lone electron pair of Bi requires. When Bi on the 0, 0, z position of the “averaged” cell is transferred to two general six-fold sites and one unoccupied twofold one of the super-cell, more structural stability is guaranteed due to compensation of its basal plane dipole momentum. Owing to the limited intensity data available, more details of the superstructure are not accessible yet. Some physical properties and solar cell application are discussed together with suggestions of ambient temperature synthesis routes of c-axis oriented nano-rod sheets.

模糊n-cell数值函数的半连续性及凸性问题的研究

利用模糊n-cell数空间上的基于模糊数的偏序关系,给出了模糊n-cell数值函数的半连续性概念,并证明了一些相关的结论。同时讨论了模糊n-cell数值函数的凸性问题,并给出了模糊n-cell数值函数在半连续条件下成为凸的充分条件。