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.

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.

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).

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.

Effect of electromagnetic force on turbulent flow of molten metal in aluminum electrolysis cells

The standard k-ε model was adopted to simulate the flow field of molten metal in three aluminum electrolysis cells with different anode risers. The Hartman number, Reynolds number and the turbulent Reynolds number of molten metal were calculated quantitatively. The turbulent Reynolds number is in the order of 103 , and Reynolds number is in the order of 104 if taking the depth of molten metal as the characteristic length. The results show that the molten metal flow is the turbulence of high Reynolds number, the turbulent Reynolds number is more appropriate than Reynolds number to be used to describe the turbulent characteristic of molten metal, and Hartman number displays very well that electromagnetic force inhibits turbulent motion of molten metal.

The role of bone marrow-derived cells in the origin of liver cancer revealed by single-cell sequencing

Objective:Epithelial cancers often originate from progenitor cells,while the origin of hepatocellular carcinoma(HCC)is still controversial.HCC,one of the deadliest cancers,is closely linked with liver injuries and chronic inflammation,which trigger massive infiltration of bone marrow-derived cells(BMDCs)during liver repair.Methods:To address the possible roles of BMDCs in HCC origination,we established a diethylnitrosamine(DEN)-induced HCC model in bone marrow transplanted mice.Immunohistochemistry and frozen tissue immunofluorescence were used to verify DENinduced HCC in the pathology of the disease.The cellular origin of DEN-induced HCC was further studied by single cell sequencing,single-cell nested PCR,and immunofluorescence-fluorescence in situ hybridization.Results:Studies by using single cell sequencing and biochemical analysis revealed that HCC cells in these mice were coming from donor mice BMDCs,and not from recipient mice.Furthermore,the copy numbers of mouse orthologs of several HCC-related genes previously reported in human HCC were also altered in our mouse model.DEN-induced HCCs exhibited a similar histological phenotype and genomic profile as human HCCs.Conclusions:These results suggested that BMDCs are an important origin of HCC,which provide important clues to HCC prevention,detection,and treatments.

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.

Single-cell DNA methylome analysis of circulating tumor cells

Objective: Previous investigations of circulating tumor cells(CTCs) have mainly focused on their genomic or transcriptomic features, leaving their epigenetic landscape relatively uncharacterized. Here, we investigated the genome-wide DNA methylome of CTCs with a view to understanding the epigenetic regulatory mechanisms underlying cancer metastasis.Methods: We evaluated single-cell DNA methylome and copy number alteration(CNA) in 196 single cells,including 107 CTCs collected from 17 cancer patients covering six different cancer types. Our single-cell bisulfite sequencing(sc BS-seq) covered on average 11.78% of all Cp G dinucleotides and accurately deduced the CNA patterns at 500 kb resolution.Results: We report distinct subclonal structures and different evolutionary histories of CTCs inferred from CNA and DNA methylation profiles. Furthermore, we demonstrate potential tumor origin classification based on the tissue-specific DNA methylation profiles of CTCs.Conclusions: Our work provides a comprehensive survey of genome-wide DNA methylome in single CTCs and reveals 5-methylcytosine(5-m C) heterogeneity in CTCs, addressing the potential epigenetic regulatory mechanisms underlying cancer metastasis and facilitating the future clinical application of CTCs.

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.

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.

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.

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.

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。

Single‑cell sequencing reveals the reproductive variations between primiparous and multiparous Hu ewes

Background In the modern sheep production systems,the reproductive performance of ewes determines the economic profitability of farming.Revealing the genetic mechanisms underlying differences in the litter size is important for the selection and breeding of highly prolific ewes.Hu sheep,a high-quality Chinese sheep breed,is known for its high fecundity and is often used as a model to study prolificacy traits.In the current study,animals were divided into two groups according to their delivery rates in three consecutive lambing seasons(namely,the high and low reproductive groups with≥3 lambs and one lamb per season,n=3,respectively).The ewes were slaughtered within 12 h of estrus,and unilateral ovarian tissues were collected and analyzed by 10×Genomics single-cell RNA sequencing.Results A total of 5 types of somatic cells were identified and corresponding expression profiles were mapped in the ovaries of each group.Noticeably,the differences in the ovary somatic cell expression profiles between the high and low reproductive groups were mainly clustered in the granulosa cells.Furthermore,four granulosa cell subtypes were identified.GeneSwitches analysis revealed that the abundance of JPH1 expression and the reduction of LOC101112291 expression could lead to different evolutionary directions of the granulosa cells.Additionally,the expression levels of FTH1 and FTL in mural granulosa cells of the highly reproductive group were significantly higher.These genes inhibit necroptosis and ferroptosis of mural granulosa cells,which helps prevent follicular atresia.Conclusions This study provides insights into the molecular mechanisms underlying the high fecundity of Hu sheep.The differences in gene expression profiles,particularly in the granulosa cells,suggest that these cells play a critical role in female prolificacy.The findings also highlight the importance of genes such as JPH1,LOC101112291,FTH1,and FTL in regulating granulosa cell function and follicular development.

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

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。

Surreal Numbers, Dyadic Rational Numbers and Cell Proliferation

The growth of multicellular organisms is directly related to cell proliferation through the cell cycle, in which a single cell grows and divides to produce two daughter cells. This process leads to an exponential increase in cell number and serves as a rapid mechanism for tissue growth. In this article, we aim to establish a connection between cell proliferation and surreal numbers, demonstrating that dyadic rational numbers emerging from surreal number theory may be related to growing cell numbers.

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.

Genomic characterization of esophageal squamous cellcarcinoma:insights from next-generation sequencing

Two major types of cancer occur in the esophagus: squamous cell carcinoma, which is associated with chronic smoking and alcohol consumption, and adenocarcinoma, which typically arises in gastric reflux-associated Barrett's esophagus. Although there is increasing incidence of esophageal adenocarcinoma in Western counties, esophageal squamous cell carcinoma(ESCC) accounts for most esophageal malignancies in East Asia, including China and Japan. Technological advances allowing for massively parallel, high-throughput next-generation sequencing(NGS) of DNA have enabled comprehensive characterization of somatic mutations in large numbers of tumor samples. Recently, several studies were published in which whole exome or whole genome sequencing was performed in ESCC tumors and compared with matched normal DNA. Mutations were validated in several genes, including in TP53, CDKN2 A, FAT1, NOTCH1, PIK3 CA, KMT2 D and NFE2L2, which had been previously implicated in ESCC. Several new recurrent alterations have also been identified in ESCC. Combining the clinicopathological characteristics of patients with information obtained from NGS studies may lead to the development of effective diagnostic and therapeutic approaches for ESCC. As this research becomes more prominent, it is important that gastroenterologist become familiar with the various NGS technologies and the results generated using these methods. In the present study, we describe recent research approaches using NGS in ESCC.

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.