Global Annotation of Small RNA and MicroRNA Mature Sequences from Developing Ovules of Gossypium hirsutum L.

The involvement of small RNAs in cotton fiber development is under explored.The objective of this work was to directly clone,annotate,and analyze small RNAs of developing ovules to reveal

Comparative analysis of SIMILAR to RCD ONE(SRO)family from tetraploid cotton species and their diploid progenitors depict their significance in cotton growth and development

Background SRO(Similar to RCD1)genes family is largely recognized for their importance in the growth,develop-ment,and in responding to environmental stresses.However,genome-wide identification and functional characteri-zation of SRO genes from cotton species have not been reported so far.Results A total of 36 SRO genes were identified from four cotton species.Phylogenetic analysis divided these genes into three groups with distinct structure.Syntenic and chromosomal distribution analysis indicated uneven distribu-tion of GaSRO,GrSRO,GhSRO,and GbSRO genes on A2,D5 genomes,Gh-At,Gh-Dt,Gb-At,and Gb-Dt subgenomes,respectively.Gene duplication analysis revealed the presence of six duplicated gene pairs among GhSRO genes.In promoter analysis,several elements responsive to the growth,development and hormones were found in GhSRO genes,implying gene induction during cotton growth and development.Several miRNAs responsive to plant growth and abiotic stress were predicted to target 12 GhSRO genes.Organ-specific expression profiling demonstrated the roles of GhSRO genes in one or more tissues.In addition,specific expression pattern of some GhSRO genes dur-ing ovule development depicted their involvement in these developmental processes.Conclusion The data presented in this report laid a foundation for understanding the classification and functions of SRO genes in cotton.

Usefulness of Newly Devised Clomiphene Citrate Administration Method Compared with the Conventional Method in Ovulation and Pregnancy

Objective: Since not all women wish to conceive a child through aggressive treatment, we investigated the usefulness of modified repeated intracyclic clomiphene citrate (CC) therapy (repeated CC therapy) as a newly devised administration method. Methods: We evaluated the effects of CC administration on menstrual cycle length and retrospectively compared ovulation and pregnancy in 220 women who received CC at our hospital. Patients in the conventional method group received 50 mg per day for five days, starting on the fifth day of menstruation (withdrawal bleeding). Groups with and without menstrual cycle shortening after conventional CC administration were compared. The repeated CC therapy group was also compared with the non-shortened group. Repeated CC therapy was administered for the first five days as in the conventional method, and a second five-day repeat treatment was administered after an interval of five to seven days. Pregnancy rates, including indirect pregnancies, were evaluated by three different methods. Results: Ovulation and pregnancy rates were significantly better in the shortened group than in the non-shortened group (P < 0.001 and P = 0.010, respectively). Even in the non-shortened group, ovulation and pregnancy rates including indirect pregnancies were significantly improved when ovulation was observed with repeated CC therapy (P < 0.001 and P = 0.022, respectively). Conclusions: For patients whose menstrual cycle was not improved or shortened, repeated CC therapy as the newly devised CC administration method is useful as the next step after the conventional CC administration method.

A Malvaceae-specific miRNA targeting the newly duplicated GaZIP1L to regulate Zn^(2+)ion transporter capacity in cotton ovules

MicroRNAs(miRNAs)play critical roles in regulating gene expression in plants,yet their functions underlying cultivated diploid Gossypium arboreum cotton ovule development are largely unknown.Here,we acquired small RNA profiles from G.arboreum ovules and fibers collected at different growth stages,and identified 46 novel miRNAs that accounted for 23.7%of all miRNAs in G.arboreum reported in the latest plant sRNA database.Through analysis of 84(including 38 conserved)differentially expressed G.arboreum miRNAs,we detected 215 putative protein-coding genes in 26 biological processes as their potential targets.A Malvaceae-specific novel miRNA named gar-miRN44 was found to likely regulate cotton ovule growth by targeting to a newly duplicated Zn^(2+)ion transporter gene GaZIP1L.During cotton ovule development,gar-miRN44 transcript level decreased sharply after 10 to 15 days post-anthesis(DPA),while that of the GaZIP1L increased significantly,with a concomitant increase of Zn^(2+)ion concentration in late ovule developmental stages.Molecular dynamics simulation and ion absorption analysis showed that GaZIP1L has stronger Zn2+ion binding ability than the original GaZIP1,indicating that the newly evolved GaZIP1L may be more suitable for maintaining high Zn2+ion transport capacity that is likely required for cotton ovule growth via enhanced cellulose synthase activities.Our systematic miRNA profiling in G.arboreum and characterization of gar-miRN44 not only contribute to the understanding of miRNA function in cotton,but also provide potential targets for plant breeding.

多囊卵巢综合征无排卵的胰岛素信号和雄激素合成的新遗传风险和代谢特征

促排卵是多囊卵巢综合征(PCOS)不孕症的一线治疗方案。卵巢对促排卵治疗的排卵应答差被认为与胰岛素抵抗和高雄激素血症相关。在一个包含1000名PCOS不孕妇女(PCOSAct)的前瞻性队列中,我们开展了一项全外显子联合靶向单核苷酸多态性(SNP)测序以及代谢组学研究。在全基因组水平找出与无排卵显著相关的常见变异和罕见突变,并通过机器学习算法构建排卵预测模型。研究发现,ZNF438基因中标记为rs2994652(p=2.47×10^(-8))的常见变异和REC114基因中的一个罕见功能突变(rs182542888,p=5.79×10^(-6))与促排卵治疗失败显著相关。携带rs2994652 A等位基因和REC114 p.Val101Leu(rs182542888)的PCOS不孕妇女进行促排卵治疗的总排卵率更低(分别为比值比(OR)=1.96,95%置信区间(CI)[1.55~2.49];OR=11.52,95%CI[3.08~43.05]),出现排卵的间隔时间更长(平均56.7天vs.49.0天,p<0.001;78.1天vs.68.6天,p=0.014)。对于rs2994652突变者,L-苯丙氨酸水平升高并与胰岛素抵抗稳态模型(HOMA-IR)指数(r=0.22,p=0.05)和空腹血糖(r=0.33,p=0.003)呈正相关;对于rs182542888突变者,花生四烯酸代谢产物水平下降并与升高的抗苗勒管激素(r=-0.51,p=0.01)和总睾酮(r=-0.71,p=0.02)呈负相关。整合基因变异位点、代谢产物及临床特征的联合预测模型可提高对排卵的预测能力[曲线下面积(AUC)=76.7%]。ZNF438基因的一个常见变异和REC114基因的一个罕见功能突变,以及与二者相关的苯丙氨酸和花生四烯酸代谢物改变,与PCOS女性不孕症的促排卵治疗失败相关。

Clinical Application of Double Ovulation Stimulation in Patients with Diminished Ovarian Reserve and Asynchronous Follicular Development Undergoing Assisted Reproduction Technology

Objective This study aimed to compare the clinical effects of double ovulation stimulation(DouStim)applied during the follicular and luteal phases with the antagonist protocol in patients with diminished ovarian reserve(DOR)and asynchronous follicular development undergoing assisted reproductive technology(ART).Methods The clinical data of patients with DOR and asynchronous follicular development receiving ART from January 2020 to December 2021 were retrospectively analyzed.The patients were divided into two groups according to their ovulation stimulation protocol:DouStim group(n=30)and antagonist group(n=62).Assisted reproduction and clinical pregnancy outcomes were compared between the two groups.Results In the DouStim group,the number of oocytes retrieved,metaphase II(MII)oocytes,two-pronuclei(2PN),day 3(D3)embryos,D3 high-quality embryos as well as blastocyst formation,implantation,and human chorionic gonadotropin-positive rates were significantly greater than those in the antagonist group(all P<0.05).No significant differences were found in MII,fertilization,or continued pregnancy rates at the first frozen embryo transfer(FET),in-vitro fertilization(IVF)cancellation,or early medical abortion rates between the groups(all P>0.05).Except for the early medical abortion rate,the DouStim group generally had favorable outcomes.In the DouStim group,the dosage and duration of gonadotropin and the fertilization rate were significantly greater in the first ovulation stimulation induction than in the second ovulation stimulation induction(P<0.05).Conclusion The DouStim protocol efficiently and economically obtained more mature oocytes and high-quality embryos for patients with DOR and asynchronous follicular development.

The sperm-interacting proteome in the bovine isthmus and ampulla during the periovulatory period

Background Spermatozoa interact with oviduct secretions before fertilization in vivo but the molecular players of this dialog and underlying dynamics remain largely unknown.Our objectives were to identify an exhaustive list of sperm-interacting proteins(SIPs)in the bovine oviduct fluid and to evaluate the impact of the oviduct anatomical region(isthmus vs.ampulla)and time relative to ovulation(pre-ovulatory vs.post-ovulatory)on SIPs number and abundance.Methods Pools of oviduct fluid(OF)from the pre-ovulatory ampulla,pre-ovulatory isthmus,post-ovulatory ampulla,and post-ovulatory isthmus in the side of ovulation were collected from the slaughterhouse.Frozen-thawed bull sperm were incubated with OF or phosphate-buffered saline(control)for 60 min at 38.5℃.After protein extraction and digestion,sperm and OF samples were analyzed by nanoLC-MS/MS and label-free protein quantification.Results A quantitative comparison between proteins identified in sperm and OF samples(2333 and 2471 proteins,respectively)allowed for the identification of 245 SIPs.The highest number(187)were found in the pre-ovulatory isthmus,i.e.,time and place of the sperm reservoir.In total,41 SIPs(17%)were differentially abundant between stages in a given region or between regions at a given stage and 76 SIPs(31%)were identified in only one region×stage condition.Functional analysis of SIPs predicted roles in cell response to stress,regulation of cell motility,fertilization,and early embryo development.Conclusion This study provides a comprehensive list of SIPs in the bovine oviduct and evidences dynamic spatiotemporal changes in sperm-oviduct interactions around ovulation time.Moreover,these data provide protein candidates to improve sperm conservation and in vitro fertilization media.

VvAGAMOUS Affect Development of Four Different Grape Species Ovary

Grape pistil has an important influence on fruit size and quality.However,there were few studies on grape ovary,and the development process of the ovary is still unclear.Therefore,in this paper,four different grape varieties with different lengths of small inflorescences,namely‘Musct Hambourg’grape(Vitis vinifera),‘Concord’grape(Vitis labrusca),‘ShanPuTao’grape(Vitis amurensis)and‘GongNiang2Hao’grape(Vitis amurensis×Vitis vinifera)were used as test materials.Four varieties ovary were significant differences by means of stereomicroscope,paraffin section.The expression of ovary determining gene VvAGAMOUS(VvAG)and its development related genes VvCRABS CLAW(VvCRC)and VvAGAMOUS-LIKE 11(VvAGL11)with similar functions during the development of different grape varieties were preliminarily explored using fluorescence quantitative test.The relationship between VvAG and VvCRC,VvAG and VvAGL11 were analyzed using Y1H assay.Our results showed that there were obvious abdominal sutures on the surface of expect for‘Musct Hambourg’grape,and existing poly carpels.The ovary development of‘ShanPuTao’and‘GongNiang2Hao’grape was completed when the inflorescence length was less than 1 cm,while the‘Concord’and‘Musct Hambourg’grape were fully developed when the length of inflorescence was 3–4 and 4–5 cm,respectively.VvAG and VvCRC began to express in large quantities after the formation of stamen primordia,while VvAGL11 during the forming of ovule primordia.Therefore,VvAG and VvCRC mainly regulated the development of stamens and carpels and also promote the development of ovules,while VvAGL11 major regulated the development of ovules.The promoters of VvCRC and VvAGL11 were bound by VvAG.This study provides an important theoretical basis for further research on the molecular mechanism of grape ovary development.

Research Progress on Mechanism of Ovulation Disorder in Polycystic Ovary Syndrome

Polycystic ovary syndrome(PCOS)is a reproductive endocrine disease characterized by ovulation disorder,hyperandrogenism and/or polycystic ovary.Follicular development disorder caused by PCOS accounts for up to 75%of anovulatory infertility,which is a common cause of infertility and affects up to 15%of reproductive aged women worldwide.Oxidative stress,chronic inflammation,endoplasmic reticulum stress and other pathological conditions coexist in the ovarian microenvironment of PCOS,which are further aggravated under the action of high levels of androgens,synergistically deteriorating the follicular microenvironment,leading to ovulation disorders in PCOS patients.This paper briefly reviewed the research on the mechanism of PCOS ovulation disorder in recent years.

利血平和丘脑下部促黄体素释放激素类似物(LHRH-A)对大鳞副泥鳅促性腺激素细胞的分泌活动和排卵的促进作用

利血平能使神经分泌细胞的儿茶酚胺贮存量减少甚至消耗殆尽,导致多巴胺作为GRIF的作用减弱,从而显著增强LHRH-A促进GtH分泌和诱导排卵的效应。对大鳞副泥鳅,利血平和LHRH-A同时注射或者利血平比LHRH-A提前3小时注射,都能激活GtH细胞,显著增强其分泌活动并诱导排卵。利血平的最低有效剂量为1—5微克/克体重。利血平(25微克/克体重)+LHRH-A(0.05微克/克体重)的催产效果和多巴胺拮抗物pimozide(1微克/克体重)+LHRH-A(0.05微克/克体重)的催产效果一样,排卵率都达到了100％。

脉冲式静脉注射促黄体激素释放激素对不孕症促排卵的疗效观察

自八十年代初开始,采用脉冲式促黄体激素释放激素(LHRH)诱导排卵,迄今已有许多成功的报道。Asch[1]作了统计,这种疗法的排卵周期率为90％,妊娠周期率为30～15％。国内于1973年人工合成了LHRH并已生产自动注射泵,脉冲式给药方法的应用在国内亦正方兴未艾。本研究采用LHRH脉冲式静脉给药方法,旨在探讨各种内分泌原因所致大孕症对LHRH脉冲治疗的反应性,为LHRH广泛用于临床提供实践基础。

New taxa of Berberis (Berberidaceae) with greenish flowers from a biodiversity hotspot in Sichuan Province, China

A new species,Berberis viridiflora X.H.Li,and a new variety,Berberis sanguinea Franch.var.viridisepala X.H.Li,L.C.Zhang & W.H.Li are described and illustrated from Baoxing County,a biodiversity hotspot located on the eastern edge of Hengduan Mountains in Sichuan Province,Southwest China,Both new taxa resemble B.sanguinea Franch.var.sanguinea,but B.viridiflora differs by the greenish flowers,and the petals being truncate,obtuse,or undulate at apex;while B.sanguinea var.viridisepala differs by the greenish or yellowish green flowers.Morphological features of the pollen grains of B.sanguinea and the two new taxa are revealed by scanning election microscope.B.sanguinea var.sanguinea displays obvious similarities with a sympatric congener,Berberis multiovula T.S.Ying in the morphology of flowers,stems and leaves,especially its ovule number varies greatly from 2 to 9,thus,B.multiovula characterized by the5-ovuled ovary is reduced to a synonymy of B.sanguinea var.sanguinea.On the basis of field surveys and study of herbarium specimens,a total of 16 species and varieties of Berberis are recorded from Baoxing County,and a key is provided.Among the 16 taxa,most phenotypic variations in Chinese Berberis can be displayed,including the habit of plants,most morphological variations of stems,branches and leaves,all types of inflorescences,all color types of flowers,and nearly all types of the shape and color of fruits.Baoxing County and its adjacent Sichuan Giant Panda Sanctuaries possibly constitute an active diversification center of Berberis in eastern Hengduan Mountains of China.

Clomiphene Citrate Induces ROS-Mediated Apoptosis in Mammalian Oocytes

The clomiphene citrate (CC), a nonsteroidal triphenylethylene compound, is a first line of medicine used for the induction of ovulation in anovulatory women worldwide. In spite of high ovulation induction with the use of CC, the pregnancy rate is much lower. Such a discrepancy could be due to the peripheral anti-estrogenic effect of CC, particularly at the level of ovary, endometrium and cervical mucus. CC induces ovulation by binding to the estrogen receptors and generates hypoestrogrnic state in hypothalamus leading to release of pituitary gonadotropins. CC may have a direct effect at the level of ovary but the molecular mechanism remains unclear. Animal studies suggest that the CC induces apoptosis in granulosa cells and results hypoestrogenic state in the ovary. Reduced estradiol 17β level in the ovary affects development and maturation of oocyte leading to oocyte apoptosis. Further, CC increases hydrogen peroxide (H2O2) level and thereby bax protein expression and DNA fragmentation in cumulus-granulosa cells as well as in oocytes. The exogenous supplementation of either estradiol 17β or melatonin reduces H2O2 level in ovary, delays meiotic cell cycle progression in oocyte and protects oocyte apoptosis. Hence, supplementation of estradiol 17β or melatonin along with CC could be beneficial to protect granulosa cell as well as oocyte apoptosis and inhibit deterioration of oocyte quality. Thus, maintenance of oocyte quality may overcome the adverse effect caused due to CC treatment during infertility management.

Comparative transcriptome analysis of the effect of different heat shock periods on the unfertilized ovule in watermelon(Citrullus lanatus)

In vitro gynogenesis is an important tool used in haploid or homozygous double-haploid plant breeding.However,because of low repeatability,embryoid induction rate and quality,the molecular mechanisms remain poorly understood.Heat shock treatment can promote the transformation of the gametophytic pathway into the sporophyte pathway,which induces the occurrence of haploid.In this study,unfertilized ovaries were heat shocked for 0 h(A0)before flowering and for 0 h(A1),4 h(A3),8 h(A5),12 h(A7),and 24 h(A8),respectively,at 37℃at the first day of the flowering stage.The ovule enlargement rate was increased from 0%at 25℃to 96.8%at 37℃(24 h treatment).Thus,we aimed to investigate the gene expression patterns in unfertilized ovules of watermelon after different periods of heat shock by using RNA-Seq technology.The results showed that compared with A3,A5,A7,and A8,the biosynthesis of amino acid,glycine,serine and threonine metabolic pathways in A1 has changed significantly.This indicated that heat shock treatment affected the synthesis and transformation of amino acids during ovule expansion.The transcriptome data suggested gene expressions of ovule growth were significantly changed by heat-specific influences.The results provide new information on the complex relationship between in vitro gynogenesis and temperature.This provides a basis for further study of the mechanism of heat shock affecting the expansion of watermelon ovule.

Randomized controlled trial of Letrozole versus Clomiphene citrate for induction of ovulation in polycystic ovarian syndrome (PCOS): A Malaysian experience

Background: The purpose of this study was to compare the effectiveness of Letrozole versus Clomiphene citrate for ovulation induction in polycystic ovarian syndrome (PCOS) with infertility. Methods: This was a prospective randomized trial involving 150 women with PCOS attending the Infertility Clinic at three hospitals in Malaysia. During the initial visit, anthropometric measurements and baseline investigations were performed. Patients were randomized to 5.0 mg Letrozole daily (75 patients) or 100 mg Clomiphene citrate daily (75 patients) from the fifth until the ninth day of menstruation. Serial transvaginal scans were performed to see the dominant follicles, endometrial thickness and number of follicles. Transvaginal scans were performed serially to look for evidence of ovulation. Results: The subjects were homogenously distributed. The difference between Letrozole and Clomiphene citrate for ovulation rate was 59 (78.7%) versus 40 (53.3%). Patients taking Letrozole exhibited a mean endometrial thickness (ET) at mid cycle of menses (Day 11-D14) of9.2 mm(SD ± 2.3) versus8.4 mm(SD ± 2.2) for patients taking Clomiphene citrate, and these differences were statistically significant

Management strategy of infertility in polycystic ovary syndrome

Polycystic ovary syndrome(PCOS)is the most common endocrine and metabolic disease in women of reproductive age.PCOS is characterized by ovulatory disruption,which can lead to infertility.Patients with PCOS are also more likely to have poor pregnancy outcomes.For obese women,lifestyle interventions are recommended first,which have general health benefits.For women who have difficulty changing their lifestyle,drugs for the treatment of obesity or bariatric surgery could be considered.Clomiphene citrate is the first-line medication after weight loss that has been utilized in the past.Letrozole is supplanting clomiphene as the best option for ovulation induction for now,particularly in patients with PCOS.Metformin can improve ovulation and pregnancy rates;however,it has minimal effects in terms of raising live birth rates.Second-line therapies include gonadotropins and laparoscopic ovary drilling.In vitro fertilization can be utilized as a third-line treatment for patients with PCOS who have failed ovulation induction therapy or have other infertility factors.In summary,to achieve fertility,patients with PCOS require standardized individualized therapy.

Heterotopic tubal pregnancy with live twin birth complicating ovulation induction assisted cycle

Heterotopic pregnancy is an uncommon clinical entity in Nigeria even though some of the risk factors are highly prevalent in the population. We report a case of heterotopic pregnancy following ovulation induction with clomiphene citrate. A favourable outcome resulting in live twin birth was recorded despite the presence of poor clinical features that affects pregnancy and foetal outcome.

Ovarian pregnancy rupture following ovulation induction and intrauterine insemination:A case report

BACKGROUND Ovarian pregnancy after assisted reproductive technology treatment has rarely been reported;ovarian pregnancy following intrauterine insemination(IUI)is even rarer,and only nine cases have previously been reported.CASE SUMMARY We report a case of ovarian pregnancy rupture after ovulation induction and IUI.The patient presented with bilateral lower abdominal pain and was referred to the emergency department.Ultrasound examination revealed ovarian pregnancy and intraperitoneal bleeding.Laparoscopy revealed an ovarian pregnancy with hemoperitoneum,which was subsequently removed.Pelvic adhesions were detected intraoperatively,which were treated immediately.The patient spontaneously conceived an intrauterine pregnancy 3 mo later,which was ongoing at the time of writing this study.CONCLUSION Close attention should be paid to any history of pelvic inflammatory disease before commencing IUI treatment,and patients with such a history should be closely followed up after IUI.Early measurement of serumβ-human chorionic gonadotropin levels and ultrasonic examination are essential for timely diagnosis of ovarian pregnancy after ovulation induction and IUI to avoid more serious complications.

New observation on seed/ovule position in the fruit of Archaeanthus and its systematic implications

1.Introduction Angiosperms are the most important group of plants in the current earth ecosystem.However,little is known about the origin of angiosperms,which has been a headache for many botanists.The core question in the origin of angiosperms is the homology of carpels.Formerly,Arber EAN and Parkin J(1907)speculated that the carpels in angiosperms were derived from the former megasporophylls bearing ovules along their margins.

Simultaneous Letrozole and Clomiphene Citrate versus Letrozole Alone in Clomiphene Citrate Resistant Polycystic Ovary Syndrome: A Randomised Controlled Trial

Background: Polycystic ovary syndrome (PCOS) is a common cause of anovulatory infertility. The therapeutic strategies for clomiphene citrate (CC)- resistant patients include the addition of corticosteroids, extended duration of clomiphene, gonadotrophin therapy, laparoscopic ovarian drilling, in vitro fertilization or the use of aromatase inhibitors recently. Letrozole decreases estrogen levels in the body, so it releases the hypothalamus and/or pituitary gland from the negative feedback of estrogen. This increases levels of gonadotrophins, which stimulates follicular growth. Objectives: To evaluate the role of letrozole alone and simultaneous use of letrozole and clomiphene citrate (CC) for ovulation induction in patients with clomiphene citrate-resistant PCOS (CCR-PCOS). Patients and Methods: This open-label randomised controlled study was conducted in the Department of Obstetrics and Gynecology, Faculty of Medicine, Zagazig University, Egypt during the period from February 2018 to June 2019. The study included 60 CCR-PCOS patients who were randomly allocated by independent personnel into two arms: group A (letrozole alone) or B (letrozole + CC). In either group, monitoring the mean follicular diameter and endometrial thickness in the days 10, 12, and 14 of the cycle by transvaginal ultrasound and Measurement of serum Progesterone (ng/ml) 7 days after the expected time of ovulation. Results: We investigated various clinical and sonographic factors that may predict the outcome of the method of induction of ovulation in CCR-PCOS with no significant affection for the results. There was a non-significant difference between the studied groups regarding ovulation and pregnancy per cycle or per patient. Conclusion: Letrozole alone or simultaneous use of letrozole and CC offers a good second-line option for induction of ovulation in CCR-PCOS patients. However, the combination of CC and letrozole did not add any benefit over the use of letrozole alone regards ovulation rate, follicular volume, endometrial thickness, pregnancy rate and live birth rate.