Sex Determination in Homo sapiens as a Multi-Step Process: Potential for Development of Variants and Sex Differences in Disease Risk

Reproduction via cis-binary mechanisms appears to have evolved fairly early in the evolution of complex organisms, and a system committed to prior to evolution of humans. While the evolution of a chromosomal-specific approach has been a successful strategy for survival of a large variety of species including humans, the fidelity of sex determination leading to 100% cis-binary outcomes is not achieved in many species, with evidence for homosexual or bisexual behaviour evident in more than 1500 species. Thus, such outcomes indicates that sex determination is a multi-step process and not a single event, and as such, could lead to the appearance of variants during the process which developed much earlier than humans. Variants could arise either due to intrinsic variation in the steps of determination, or also be influenced by environmental factors of a biological or psychological nature. In contrast to homosexual variants which do not require interventions such as hormone therapy or surgery, expression of gender dysphoria, is more based in psychology, but also has biological underpinnings and can be influenced by such hormonal interventions and surgery. While the numbers of those with gender dysphoria is small (~0.6% - 1.0% of population), the attention given to this issue raises the possibility of biological and psychological environmental factors impacting the emergence of some of those expressing gender dysphoria. Furthermore, transitioning from male-to-female or female-to-male can have consequences regarding disease risks latter in life, including the appearance of autoimmune diseases. This review will attempt to review some of the evidence regarding sex determination, discuss why the system has potentially not been improved upon during evolution, how a potential role for sex chromosome function on neurodevelopment may be central to variation in humans, and how commitment to the current strategy is likely integrated into other sex-related events such as puberty, pregnancy, and menopause to ensure species survival. It will also discuss how variants in sex determination could contribute to sex differences in disease risk and how epigenetic modifications could play a role in such risk. .

Silencing transformer and transformer-2 in Zeugodacus cucurbitae causes defective sex determination with inviability of most pseudomales

transformer is a switch gene for sex determination in many insects, which cooperates with transformer-2 that is expressed in both sexes to regulate female differentiation, particularly in dipterans. Zeugodacus cucurbitae(Coquillett) is a very destructive pest worldwide, however, its sex determination pathway remains largely uncharacterized. Here, we show that the female sex ratio is sharply reduced with knockdown of either transformer or transformer-2 by RNA interference in early embryos of Z. cucurbitae. Most of the males grown from the embryos with transient transformer and transformer-2 suppression mated with wild-type females and produced mixed sex progeny, with one exception that produced only female progeny, and all of the few remaining males failed to mate with wild-type females and produced no progeny. The exceptional male and those males with mating failure were XX pseudomales as determined by the detection of Y chromosome-linked Maleness-on-the-Y, indicating that most XX pseudomales are not viable. The phenotypes of transformer and transformer-2 suggest that they play a key role in regulating sex determination and are required for female sexual development of Z. cucurbitae. Our results will be beneficial to the understanding of sex determination in Z. cucurbitae and can facilitate the development of genetic sexing strains for its biological control.

Investigation of sex determination in starry flounder(Platichthys stellatus)reveals sex chromosome evolution in Pleuronectiformes and identifies a sex-specific marker

The identification of sex chromosomes is fundamental for exploring the mechanism and evolution of sex determination.Platichthys stellatus,a species exhibiting clear sexual dimorphism and homomorphic chromosome pairs,has received limited research concerning its sex determination mechanisms.Clarifying the sex chromosome of P.stellatus will enhance our understanding of sex chromosome evolution in Pleuronectiformes.This study employed whole-genome resequencing to investigate the sex chromosome and sex determination system in P.stellatus.Notably,Chr23 was identified as the sex chromosome in P.stellatus,with the sex-determining region(SDR)occupying 48.1%of the chromosome and featuring an XX/XY system.Sex chromosome turnover was observed within Pleuronectiformes,with P.stellatus,Verasper variegatus,and Hippoglossus hippoglossus sharing a common ancestral karyotype.No inversions were detected within the SDR of P.stellatus,although chromosomal rearrangements between sex chromosomes and autosomes were identified.Additionally,a sex-specific marker for P.stellatus was ascertained,enabling genetic sex identification,with significant implications for improving breeding programs and aquaculture practices.

Determination of Chlorogenic Acid,Geniposide,Total Flavonoids and Total Triterpenes in Wulan-13

[Objectives]This study was conducted to establish a method for the determination of chlorogenic acid,geniposide,total flavonoids and total triterpenes in Wulan-13.[Methods]The contents of chlorogenic acid and geniposide were determined by HPLC,and the contents of total flavonoids and total triterpenes were determined by an ultraviolet spectrophotometer.[Results]There was a good linear relation between the mass of chlorogenic acid reference substance and the peak area in the range of 0.05-0.45μg,and the regression equation was Y=2524.1X+3.1943,(r=0.9998).A good linear relationship was found between the mass of gardenoside reference substance and the peak area in the range of 0.776-6.984μg,and the regression equation was Y=1670.5X+64.804,(r=0.9998).There was also a good linear relation between the mass of rutin reference substance and its absorbance in the range of 0.00808-0.04848 mg,and the regression equation was Y=12.916X+0.014,(r=0.999).The mass of oleanolic acid reference substance had a good linear relation with its absorbance in the range of 0.00418-0.0209 mg,and the regression equation was Y=51.89X-0.0839,(r=0.9991).[Conclusions]The content determination method is simple,reliable and reproducible,and suitable for controlling the contents of chlorogenic acid,geniposide,total flavonoids and total triterpenes in Wulan-13.

Overexpression of Kdm6b induces testicular differentiation in a temperature-dependent sex determination system

In reptiles,such as the red-eared slider turtle(Trachemys scripta elegans),gonadal sex determination is highly dependent on the environmental temperature during embryonic stages.This complex process,which leads to differentiation into either testes or ovaries,is governed by the finely tuned expression of upstream genes,notably the testis-promoting gene Dmrt1 and the ovary-promoting gene Foxl2.Recent studies have identified epigenetic regulation as a crucial factor in testis development,with the H3K27me3 demethylase KDM6B being essential for Dmrt1 expression in T.s.elegans.However,whether KDM6B alone can induce testicular differentiation remains unclear.In this study,we found that overexpression of Kdm6b in T.s.elegans embryos induced the male development pathway,accompanied by a rapid increase in the gonadal expression of Dmrt1 at 31°C,a temperature typically resulting in female development.Notably,this sex reversal could be entirely rescued by Dmrt1 knockdown.These findings demonstrate that Kdm6b is sufficient for commitment to the male pathway,underscoring its role as a critical epigenetic regulator in the sex determination of the red-eared slider turtle.

Frequentist and Bayesian Sample Size Determination for Single-Arm Clinical Trials Based on a Binary Response Variable: A Shiny App to Implement Exact Methods

Sample size determination typically relies on a power analysis based on a frequentist conditional approach. This latter can be seen as a particular case of the two-priors approach, which allows to build four distinct power functions to select the optimal sample size. We revise this approach when the focus is on testing a single binomial proportion. We consider exact methods and introduce a conservative criterion to account for the typical non-monotonic behavior of the power functions, when dealing with discrete data. The main purpose of this paper is to present a Shiny App providing a user-friendly, interactive tool to apply these criteria. The app also provides specific tools to elicit the analysis and the design prior distributions, which are the core of the two-priors approach.

Optimal urban EV charging station site selection and capacity determination considering comprehensive benefits of vehicle-station-grid

This paper presents an optimization model for the location and capacity of electric vehicle(EV)charging stations.The model takes the multiple factors of the“vehicle-station-grid”system into account.Then,ArcScene is used to couple the road and power grid models and ensure that the coupling system is strictly under the goal of minimizing the total social cost,which includes the operator cost,user charging cost,and power grid loss.An immune particle swarm optimization algorithm(IPSOA)is proposed in this paper to obtain the optimal coupling strategy.The simulation results show that the algorithm has good convergence and performs well in solving multi-modal problems.It also balances the interests of users,operators,and the power grid.Compared with other schemes,the grid loss cost is reduced by 11.1%and 17.8%,and the total social cost decreases by 9.96%and 3.22%.

On simulation of precise orbit determination of HY-2 with centimeter precision based on satellite-borne GPS technique

The HY-2 satellite carrying a satellite-borne GPS receiver is the first Chinese radar altimeter satellite, whose radial orbit determination precision must reach the centimeter level. Now HY-2 is in the test phase so that the observations are not openly released. In order to study the precise orbit determination precision and procedure for HY-2 based on the satellite- borne GPS technique, the satellite-borne GPS data are simulated in this paper. The HY-2 satellite-borne GPS antenna can receive at least seven GPS satellites each epoch, which can validate the GPS receiver and antenna design. What＇s more, the precise orbit determination processing flow is given and precise orbit determination experiments are conducted using the HY-2-borne GPS data with both the reduced-dynamic method and the kinematic geometry method. With the 1 and 3 mm phase data random errors, the radial orbit determination precision can achieve the centimeter level using these two methods and the kinematic orbit accuracy is slightly lower than that of the reduced-dynamic orbit. The earth gravity field model is an important factor which seriously affects the precise orbit determination of altimeter satellites. The reduced-dynamic orbit determination experiments are made with different earth gravity field models, such as EIGEN2, EGM96, TEG4, and GEMT3. Using a large number of high precision satellite-bome GPS data, the HY-2 precise orbit determination can reach the centimeter level with commonly used earth gravity field models up to above 50 degrees and orders.

Determination of 1-Phenylpropanol in Raw Material and Preparations by High-Performance Liquid Chromatograph

Ahn To establish an RP-HPLC method for determination of content of 1-phenylpropanol in its raw material and preparations. Methods Chromatography was carried out on a Dikma DiamonsilTM ODS column, using a mobile phase of methanol-water （55：45） with a flow rate at 1.0 mL·min^-1. The detection wavelength was 258 nm. Results Under the chromatographic condition, the peaks of 1-phenylpropanol and its related impurities separated completely; noninterference between the principal agent and adjuvants in preparations was performed. The calibration curve was linear over the range of 250 - 750μg·mL^-1 with the correlation coefficient of 0. 999 9. The average recovery was 100.2% （ RSD = 1.35% ）. Conclusion This method is simple, rapid, accurate, specific, and can be used to detect the content and related compounds of phenylpropanol in its raw material and preparations.

Construction of universal quantitative models for the determination of cefoperazone sodium/sulbactam sodium for injection from different manufacturers using near-infrared reflectance spectroscopy

To develop near-infrared （NIR） reflectance spectroscopic methods for the quantitative analysis of cefoperazone sodium/ sulbactam sodium from different manufacturers for injection powder medicaments. Various powders of cefoperazone sodium/ sulbactam sodium were directly analyzed by non-destructive NIR reflectance spectroscopy using the spectrometer EQUINOX55. Two quantitative methods via integrating sphere （IS） and fiberoptic probe （FOP） models were explored from 6 batches of commercial samples and 42 batches of laboratory samples at a content ranging from 30% to 70% for cefoperazone and 60% to 20% for sulbactam. The root mean square errors of cross validation （RMSECV） and the root mean square errors of prediction （RMSEP） of IS were 1.79% and 2.85%, respectively, for cefoperazone sodium, and were 1.86% and 3.08%, respectively, for sulbactam sodium; and those of FOP were 2.93% and 2.92%, respectively, for cefoperazone sodium, and were 2.23% and 3.01%, respectively, for sulbactam sodium. Based on the ICH guidelines and Ref. 12, the quantitative models were then evaluated in terms of specificity, linearity, accuracy, precision, robustness and model transferability. The non-destructive quantitative NIR methods used in this study are applicable for rapid analysis of injectable powdered drugs from different manufacturers.

Determination of β-sitosterol Content in Ethnodrug Guoshangye

[Objective] The contents of β-sitosteml in Guoshangye from different habi- tats were determined. [Metlmd] UPLC-ELSD was adopted to determine the contents of β-sitosteml in Guoshangye from different habitats, in which BEH C18 column （2.1 mmx50 ram, 1,7 urn） was used with a mobile phase of methanol-water at a flow rate was of 0.3 ml/min, and the column temperature was of 30 ℃; and an evaporative light-scattering detector was used, the temperature in drift tube was of 90 ℃ and the pressure of N2 was of 20 psi. [Concludoa] This research provided theoretical foundation for the determination of β-sitosterol in Ethnodrug Guoshangye.

Research into Uncertainty in Measurement of Seawater Chemical Oxygen Demand by Potassium Iodide-Alkaline Potassium Permanganate Determination Method

Using the glucose and L-glutamic-acid to prepare the standard substance according to the ratio of 1：1, and the artificial seawater and the standard substance to prepare a series of standard solutions, the distribution pattern of uncertainty in measurement of seawater COD is obtained based on the measured results of the series of standard solutions by the potassium iodide-alkaline potassium permanganate determination method. The distribution pattern is as follows： Uncertainty in measurement is big and not constant at the high end, but small and constant at the low end.

Improved mean-shift-based pitch determination

The underlying principle of pitch determination based on the mean shift algorithm is studied, and the cause of pitch error propagation in the original pseudo code is analyzed. The problem of error propagation is solved by choosing an appropriate initial pitch candidate F00. The theoretical choice guideline in a pitch epoch is obtained as ensuring the true pitch F0 satisfying F00/2 〈 F0 〈 3F00/2. The validity of the choice guideline is verified by the F00 experiment. Meanwhile, the algorithm is extended to the pitch determination in the noisy case and compared with the method of subharmonic-to-harmonic ratio （SHR）. The experimental results show that the improved algorithm bears comparison with SHR and it runs much faster than SHR.

基于“微生物-肠-脑轴”探讨补肾通腑方对APP/PS1小鼠肠道菌群及LPS/TLR4/NF-κB通路的影响

目的探讨补肾通腑方调控肠道菌群,改善阿尔茨海默病(Alzheimer’s disease,AD)模型小鼠学习记忆能力的作用机制。方法以APP/PS1小鼠为研究对象,给予补肾通腑方治疗8周,采用Morris水迷宫法观察小鼠空间学习记忆能力变化;16S rDNA检测小鼠肠道菌群丰度、多样性变化;HE染色观察海马病理形态学变化;免疫荧光检测海马区小胶质细胞活化情况;Western blot检测TLR4、NF-κB、IL-6等炎症因子表达。结果与模型组相比,补肾通腑方可以缩短AD模型小鼠逃避潜伏期、游泳路径,增加跨越平台次数(P<0.05),提升肠道菌群多样性,调节肠道菌群丰度,促进海马神经元细胞损伤修复,降低iNOS/Iba1共表达,提高Arg1/Iba1共表达(P<0.01),促进小胶质细胞从M1型向M2型转化,下调TLR4、NF-κB、IL-6等促炎因子的表达。结论补肾通腑方改善AD模型小鼠学习记忆能力的作用机制可能与调节肠道菌群介导的LPS/TLR4/NF-κB通路,从而抑制促炎型小胶质细胞活化、减轻中枢神经系统炎症、改善海马区神经元细胞损伤有关。

Orbit Determination Using Satellite-to-Satellite Tracking Data

Satellite-to-Satellite tricking (SST) data can be used to determine the orbits of spacecraft in two ways. One is combined orbit determination, which combines SST data with ground-based tracking data and exploits the enhanced tracking geometry. The other is the autonomous orbit determination, which uses only SST. The latter only fits some particular circumstances since it suffers the rank defect problem in other circumstances. The proof of this statement is presented. The nature of the problem is also investigated in order to find an effective solution. Several. methods of solution are discussed. The feasibility of the methods is demonstrated by their application to a simulation.

Simultaneous spectrophotometric determination of cobalt,copper and nickel using 1-(2-thiazolylazo)-2-naphthol by chemometrics methods

The simultaneous determination of cobalt, copper and nickel using 1-（2-thiazolylazo）-2-naphthol （first figure of this article） by spectrophotometric method is a difficult problem in analytical chemistry, due to spectral interferences. By multivariate calibration methods, such as partial least squares （PLS） regression, it is possible to obtain a model adjusted to the concentration values of the mixtures used in the calibration range. Orthogonal signal correction （OSC） is a preprocessing technique used for removing the information unrelated to the target variables based on constrained principal component analysis. OSC is a suitable preprocessing method for PLS calibration of mixtures without loss of prediction capacity using spectrophotometric method. In this study, the calibration model is based on absorption spectra in the 550-750-nm range for 21 different mixtures of cobalt, copper and nickel. Calibration matrices were formed from samples containing 0.05-1.05, 0.05-1.30 and 0.05-0.80 μg·mL^-1 for cobalt, copper and nickel, respectively. The root mean square error of prediction （RMSEP） for cobalt, copper and nickel with OSC and without OSC were 0.007, 0.008, 0.011 and 0.031,0.037, 0.032 μg· mL^-1, respectively. This procedure allows the simultaneous determination of cobalt, copper and nickel in synthetic and real samples and good reliability of the determination was proved.

需求不确定下的两阶段应急物流选址-路径研究

针对灾后应急救援在需求不确定和资源受限方面的问题,以多级应急物流网络为背景,构建了一个需求不确定下的两阶段应急选址-路径规划模型。该模型以总成本最小和救援车辆运输总距离最短为目标,采用三角模糊数刻画受灾点的不确定需求,并采用基于可信性的模糊机会约束规划方法,以消除约束条件中的不确定参数。模型第一阶段调用Gurobi求解器,求解得到应急配送中心选址结果和对受灾点的分配方案;第二阶段将选址及分配结果作为输入进行路径规划,并提出一种改进的自适应遗传算法(IAGA)对算例进行求解。然后采用自适应遗传算法(AGA)与之对比,并进行灵敏度分析。结果表明:IAGA在目标值、收敛速度和运行时间等方面均优于AGA,证明了IAGA具有一定的可行性和有效性,且可以为决策者提供较优的应急选址-路径规划方案,从而提升灾后救援的效率。

SPME-Arrow/GC-MS结合多元统计分析研究不同批次爆珠香精组分差异

以箭型固相微萃取(SPME-Arrow)为样品萃取手段,通过优化萃取头种类、氯化钠添加量、萃取温度、平衡时间及萃取时间等条件,构建了高效鉴定爆珠香精中挥发性及半挥发性化合物的箭型固相微萃取结合气相色谱-质谱联用(SPME-Arrow/GC-MS)法。采用该方法对不同批次爆珠香精进行分析,结合主成分分析(PCA)、正交偏最小二乘-判别分析(OPLS-DA)及显著性F检验等多元统计分析手段,筛选显著性差异成分。结果表明:最佳萃取条件为优选DVB/CAR/PDMS萃取头,添加2.0 g氯化钠,在60℃下平衡50 min,萃取时间为40 min;经谱库检索结合保留指数辅助定性,共鉴定出96种挥发性、半挥发性组分,目标物峰面积的日间相对标准偏差(RSD)小于10%的色谱峰数量占总峰数量的92.7%,表明方法的重复性较好;从不同批次爆珠香精中共筛选出17个潜在差异化合物,通过显著性检验,确定10种显著性差异化合物,分别为α-蒎烯、柠檬烯、桉叶油醇、异胡薄荷醇、薄荷酮(含异构体)、新异薄荷醇、乙酸新薄荷酯、三辛酸甘油酯和二辛酸单癸酸甘油酯。该法可有效区分不同批次爆珠样品的组分差异,具有客观真实、准确可靠、可视化强等特点,能够为爆珠产品质量检验提供技术支持。

法舒地尔缓解β-淀粉样蛋白1-42诱导的SH-SY5Y细胞凋亡

背景:法舒地尔对阿尔茨海默病小鼠脑内的线粒体动力学有调节作用,并且可以抑制神经炎症,但能否调节线粒体自噬和NLRP3炎症小体进而减轻β-淀粉样蛋白毒性尚不清楚。目的:探究法舒地尔对β-淀粉样蛋白1-42诱导的人源神经母细胞瘤细胞株SH-SY5Y凋亡和线粒体自噬以及NLRP3炎症小体的调节作用。方法:将SH-SY5Y细胞接种于孔板内,细胞贴壁后分3组干预:对照组不加入任何药物,模型组加入20μmol/L β-淀粉样蛋白1-42,法舒地尔组同时加入20μmol/L β-淀粉样蛋白1-42与15 mg/L法舒地尔,干预24 h后,采用MTT法检测细胞活性,TUNEL染色检测细胞凋亡,qRT-PCR和Western blot检测凋亡相关蛋白表达,免疫荧光染色和Western blot检测线粒体自噬相关蛋白表达,免疫荧光染色和Western blot检测NLRP3炎症小体相关蛋白表达。结果与结论:(1)与对照组比较,模型组细胞活性降低、凋亡率升高(P<0.05);与模型组比较,法舒地尔组细胞活性升高、凋亡率降低(P<0.05);(2)qRT-PCR和Western blot检测结果显示,与对照组比较,模型组细胞Bax mRNA与蛋白表达升高(P<0.05),Bcl-2 mRNA与蛋白表达降低(P<0.05);与模型组比较,法舒地尔组细胞Bax mRNA与蛋白表达降低(P<0.05),Bcl-2 mRNA与蛋白表达升高(P<0.05);(3)免疫荧光染色和Western blot检测结果显示,与对照组相比,模型组细胞PINK1、帕金森病蛋白和LC3蛋白表达降低(P<0.05),p62蛋白表达升高(P<0.05);与模型组相比,法舒地尔组细胞PINK1、帕金森病蛋白和LC3蛋白表达升高(P<0.05),p62蛋白表达降低(P<0.05);(4)免疫荧光染色和Western blot检测结果显示,与对照组相比,模型组细胞NLRP3、ASC、Caspase-1和白细胞介素1β蛋白表达升高(P<0.05);与模型组相比,法舒地尔组细胞NLRP3、ASC、Caspase-1和白细胞介素1β蛋白表达降低(P<0.05);(5)结果表明,法舒地尔可以减轻β-淀粉样蛋白1-42诱导的SH-SY5Y细胞凋亡,其机制可能与激活线粒体自噬且抑制NLRP3炎症小体激活有关。

Formation of multiple trimethylsilyl derivatives in the derivatization of 17α-ethinylestradiol with BSTFA or MSTFA followed by gas chromatography-mass spectrometry determination

N,O-bis（trimethylsily）trifluoroacetamide （BSTFA） and N-methyl-N（trimethylsily） trifluoroacetamide （MSTFA） are common derivatization reagents used in the GC-MS analysis of estrogen steroids such as estrone （El） and 17α-ethinylestradiol （EE2）. In this study, three trimethylsilyl （TMS） steroid derivatives, mono- and di-trimethylsilyl EE2 and mono-trimethylsilyl El, were observed during the derivatization of EE2 with BSTFA or MSTFA and/or GC separation. Factors influencing the production of multiple TMS derivatives and their relative abundance were examined. It was found that both methanol and bisphenol A competed with estrogenic esteroids when reacting with silylation reagents, and thus affected the formation of TMS derivatives and their relative abundance in the derivatization products. Methanol was found to be more reactive than bisphenol A with the BSTFA reagent. None of the three solvents tested in this study could prevent the generation of multiple TMS derivatives during the derivatization of EE2 with BSTFA, followed by GC analysis. A similar result was observed using MSTFA as the derivative reagent followed by GC analysis. Thus, the suitability of BSTFA or MSTFA as the derivatization reagent for the determination of E1 and EE2 by GC-MS, under the conditions reported here, is questionable. This problem can be solved by adding trimethylsilylimidaz （TMSI） in the BSTFA reagent as recommended, and the performance of the method has been proved in this study.