DATA REPORT article Front. Reprod. Health, 11 July 2022Sec. Assisted Reproduction https://doi.org/10.3389/frph.2022.886277

Background: Diminished ovarian reserve (DOR) contributes significantly to female infertility. Embryo quality affects pregnancy outcome. However, the factors that influence pregnancy outcomes in DOR patients are not well established. In this study, we analyzed the medical records of infertile women with DOR who underwent ART to reveal the possible correlation between clinical characteristics and embryo quality. Methods: A retrospective cohort study was conducted using 3390 medical record data on IVF cycles from Dalian Municipal Women and Children’s Medical Center (Group) from January 2015 to December 2020. The data was divided into poor-quality embryos group and high-quality embryos group according to the embryo status. We summarized the demographic and clinical characteristics (age, infertility years, infertility type, AFC, and BMI), medical history, reproductive hormone parameters (AMH, FSH, LH, PRL, and E 2 ), and carbohydrate and lipid metabolic indexes (CHO, TG, LP, LDL, HDL, FINS, Glucose, and Hcy). To further evaluate the Hcy levels in patients with different embryo quality, we performe subgroup analysis. In order to further evaluate the Hcy levels of patients with different embryo quality, we conducted a subgroup analysis, which was divided into three subgroups: Hcy ≥15, 15> Hcy >8 and Hcy ≤8. A total of 1023patients with DOR were included in this study. Results: Of these, 483 cases were with high-quality embryos (the number of high quality embryo, number≥1), and 540 cases with poor-quality embryos (no high quality embryo, number=0). Demographic characteristics of the two groups were similar. However, the level of serum Hcy was significantly higher in the poor-quality embryo group compared with the high-quality embryo group (P < 0.05). In addition, the levels of serum AMH and PRL were statistically different between the two groups. Patients with serum Hcy ≥ 15 μmol/L were at higher risk of having poor-quality embryos (82.1%) compared to patients with serum Hcy less than 8 μmol/L (43.9%) and at 8~15 μmol/L (48.1%) (c2 =15.859, P < 0.001). Conclusion: The presented study demonstrated that high serum Hcy level is linked to poor-quality embryos. Moreover, Hcy coupled with age might be a sensitive marker for embryo quality after IVF.

目的 观察坤灵丸对卵巢储备功能下降(DOR)患者体外受精-胚胎移植(IVF/ICSI-ET)胚胎质量的影响.方法 将2020年1月至2021年4月于本中心行IVF/ICSI-ET助孕治疗的DOR患者随机分为两组,坤灵丸组(n=50)和对照组(n=50).对两组血清雌二醇(E2)、卵泡刺激素(FSH)、黄体生成素(LH)激素水平及窦卵泡(AFC)进行比较.对DOR患者IVF-ET采卵后实验室指标,包括获卵数目、2原核(2PN)受精数目、可利用胚胎数目、优质胚胎数目及优质胚胎率、移植一次妊娠结局、绒毛膜促性腺激素日(即hCG日)雌激素(E2)浓度进行统计分析.结果 坤灵丸组用药后FSH和LH水平低于用药前(P<0.05);坤灵丸组优质胚胎数目和优质胚胎率显著高于对照组(P<0.05);2PN受精数目、AFC、hCG日E2浓度两组差异无统计学意义;坤灵丸组获卵数目、可利用胚胎数目、移植一次妊娠率略高于对照组,但差异无统计学意义.结论 坤灵丸能有效调整DOR患者激素水平,改善DOR患者体外受精-胚胎移植的胚胎质量,有望作为DOR患者接受体外受精-胚胎移植前的干预药物开展更深入的临床观察.

目的 构建科学合理的组织学与胚胎学形成性评价体系,并探索其对中医学生自主学习能力和学习效果的影响.方法 选取该校中医学专业2015级、2017级和2018级五年制学生作为研究对象,其中2015级76名,2017级66名,2018级68名.对体现学生理解与应用能力的论述题成绩进行分析,采用调查问卷评价形成性评价对学生自主学习能力的影响.结果 2017级、2018级80～100分数段比例明显提高,而<60分比例明显降低.2017级、2018级学生论述题得分均高于2015级学生,差异有统计学意义(P<0.05).在学习意识方面,95.38％(124/130)的学生认为形成性评价有助于激发学习兴趣,自觉获取学习资源和专业前沿信息,其中2017级中31.82％(21/66)的学生在科研项目方面参与度不足.在学习规划方面,85.38％(111/130)的学生认为形成性评价有助于制定学习目标和学习计划,合理安排学习时间.在学习实施方面,80.00％(104/130)的学生认为形成性评价有助于自我调节学习态度,将理论与临床相结合.在学习反馈方面,91.54％(119/130)的学生认为形成性评价有助于每学期结束后的思考与反思,并及时调整学习计划.结论 在组织学与胚胎学教学中,应用形成性评价体系可明显提高学生自主学习能力.

针对解剖学、组织学与胚胎学和病理学的实验附属于各自的理论课,在不同的学期授课,各门实验课之间缺乏关联,造成知识的割裂的现状,梳理现有形态学实验课程,在教材编写、课程设立、教学手段方面进行了改革探索.尝试建立以人体系统、器官为主线,以疾病为中心,从正常器官结构到异常器官结构,从整体器官形态观察到镜下微细结构的实验模式.希望不断深入形态学实验教学改革,一方面有利于学生对关联知识的融会贯通,另一方面培养学生的临床意识.

目的 运用网络药理学的研究方法分析紫草治疗乳腺癌的作用机制,并通过人乳腺癌MDA-MB-231细胞进行实验验证.方法 通过中药系统药理数据库和分析平台(TCMSP)等数据库筛选紫草的活性成分和治疗乳腺癌的作用靶点,并进行基因本体(GO)富集及京都百科全书和基因组(KEGG)通路富集分析.选择紫草关键活性成分紫草素对MDA-MB-231细胞进行实验验证,将细胞分为空白组、紫草素,通过细胞增殖实验测定紫草素对MDA-MB-231细胞增殖的影响,并进行线性拟合计算,得出细胞生长抑制率为0％,25％和50％时的药物浓度分别为0.3,1.6和2.7 mol·L-1,将其作为蛋白质印迹法的处理浓度,检测B淋巴细胞瘤-2蛋白(Bcl-2)等靶点蛋白[如磷脂酰肌醇3-激酶(PI3K)和蛋白激酶B(Akt)]的表达.结果 紫草素0.62,1.25,2.50和5.00 mol·L-1组的细胞抑制率分别为1.32％,21.24％,50.17％和72.37％,与空白组相比,紫草素1.25,2.50,5.00 mol·L-1组差异均有统计学意义(均P<0.01).空白组和低、中、高3个浓度(0.3,1.6,2.7 mol·L-1)紫草素组的PI3K的蛋白相对表达量分别为0.76±0.17,0.86±0.21,0.71±0.25和0.42±0.04;这4组Akt蛋白相对表达量分别为1.15±0.17,1.08±0.15,1.09±0.05和0.73±0.11;这4组Bcl-2蛋白相对表达量分别为2.25±0.30,2.02±0.14,1.64±0.16和1.24±0.20.与空白组相比,2.7 mol·L-1紫草素组能够降低PI3K、Akt蛋白表达并显著下调Bcl-2蛋白表达;1.6 mol·L-1紫草素组能够下调Bcl-2蛋白表达,以上差异均有统计学意义(P<0.05,P<0.01).结论 紫草素能够下调Bcl-2、PI3K、Akt蛋白表达,抑制人乳腺癌MDA-MB-231细胞增殖.

目的 探讨紫草素(shikonin,SK)在雌激素作用下对人乳腺癌MCF-7细胞周期蛋白(cyclin D1)表达的影响.方法 体外培养人乳腺癌MCF-7细胞,不同浓度的紫草素及在雌激素干预下作用于细胞后,通过免疫组织化学技术、Western blot法检测cyclin D1蛋白的表达变化.结果 与空白对照组比较,雌二醇组cyclin D1表达量显著增多,SK各组表达含量显著降低,且cyclin D1蛋白表达量随SK浓度升高逐渐被抑制(P＜0.05);雌二醇干预后,SK对cyclin D1蛋白量的表达同样具有抑制作用(P＜0.05).结论 SK能降低MCF-7细胞中cyclin D1蛋白水平表达,具有浓度依赖性,并且对雌激素上调cyclin D1表达具有明显的抑制作用.

大学阶段是大学生知识体系搭建和价值观塑造成型的关键时期,在全员育人、全程育人、全方位育人德育教育新模式下,课程思政作为育人新理念,是推进"三全育人"的重要环节和体现.结合北京中医药大学组织学与胚胎学教学实际,论述了实施课程思政的优势和必要性,对目前思政教学存在的问题进行了分析,并对实施课程思政的有效措施进行了探讨.

目的:应用网络药理学方法,从双黄连方剂中预测具有抗肿瘤活性的组分中药,并通过离体细胞实验与动物实验对其抗CT26结肠癌活性进行初步验证.方法:基于中药系统药理学数据库(Traditional Chinese Medicine Systems Pharmacology Database,TCMSP),《中药活性成分分析手册》,药品生物信息学和化学信息学数据库(DrugBank),信号通路数据库(Reactome)及人类蛋白质参考数据库(Human protein reference database,HPRD),构建蛋白-蛋白相互作用网络(proteinprotein interactions network,PPI)和化合物靶标网络,挖掘化合物的药理作用,发现与PPI网络节点距离较短的化合物,确定组成中药的配伍.以CT26结肠癌细胞为实验材料,结合离体和在体实验初步验证组分中药的抗肿瘤作用.结果:从双黄连组分中预测并发现“黄芩苷、连翘酯苷A和绿原酸”3个化合物配伍具有潜在抗肿瘤活性.验证结果表明,细胞实验中,与空白组比较,双黄连组分中药能显著抑制CT26细胞的增殖和迁移(P＜0.01);动物实验中,与荷瘤模型组比较,双黄连组分中药治疗组小鼠皮下肿瘤体积增长缓慢、肿瘤重量显著降低(P＜0.01).结论:结合数据库挖掘和化合物靶点网络构建、分析等方法从双黄连方剂中发现具有抗肿瘤活性的组分中药“黄芩苷、连翘酯苷A和绿原酸”.经验证,该组分中药具有较好的抗CT26结肠癌活性.

目的 观察二仙汤对卵巢早衰(premature ovarian failure,POF)大鼠的抗氧化应激效应.方法 雌性SD大鼠40只,监测一周动情周期,筛选有正常动情周期的大鼠入组,随机分为4组:正常组、模型组、阳性药组和二仙汤组.除正常组外,其余3组使用顺铂注射液造POF模型,阳性药组和二仙汤组在造模同时进行药物干预,灌胃期间监测动情周期,15 d后麻醉取材,应用放免法检测血清性激素卵泡刺激素(follicle timulating hormone,FSH)、黄体生成素(luteinizing hormone,LH)及雌激素(estrogen,E2)的水平以及氧化应激相关指标丙二醛(malondialdehyde,MDA)、抗氧化指标超氧化物歧化酶(superoxide dismutase,SOD)和过氧化氢酶(catalase,CAT)的含量,显微镜下观察各组卵巢组织的病理改变.结果 与正常组相比较,模型组大鼠动情周期及动情间期明显延长(P<0.01),血清FSH、LH、MDA含量显著升高(P<0.01),E2、SOD及CAT水平显著降低(P<0.01),卵巢组织中闭锁卵泡增多,窦卵泡数减少.与模型组相比,阳性药组与二仙汤组动情周期缩短(P<0.05或P<0.01),血清FSH、LH、MDA含量显著降低(P<0.05或P<0.01),E2、SOD及CAT的含量显著上升(P<0.05或P<0.01),闭锁卵泡数量减少,窦卵泡数增多.结论 二仙汤能通过调节POF大鼠的的抗氧化能力来改善卵巢功能,减少氧化应激带来的损伤从而治疗POF.

牛建昭教授从事妇科临床、教学、科研近50年,擅长治疗各种妇科疑难杂症,其经验丰富且独到,尤其在治疗子宫内膜异位症方面亦颇有建树.子宫内膜异位症是由于各种病理因素导致有活性的内膜组织(包括腺体和间质),不仅生长在宫腔而且还附着在子宫内膜以外的其他部位,这些异常生长的内膜组织会引起女性经期痛经,严重者可引发不孕.中医学认为子宫内膜异位症的基本病机是"瘀阻胞宫冲任",牛建昭教授在辨证论治、四诊合参的基础上,结合患者临床症状,认为青春期子宫内膜异位症的核心病机为"肾虚瘀结",在治疗时注重"西医辨病,中医分型,中西结合,个性化治疗"的诊治思路,根据月经周期遣方用药,强调审因论治,注重情志调节.此外,对青春期女性确立了调经促卵、理气止痛、温补肾阳的治疗原则.根据患者个体差异随症加减,临床疗效显著.

Estrogen receptor (ER) is expressed in most Breast cancer (BC) patients. G protein-coupled estrogen receptor (GPER), which is a membrane-bound estrogen receptor, is associated with the tumor development and progression in BC. Shikonin (SK) is a natural compound that is known to have anti-tumor effects. This study aims to assess the effects of shikonin on the cell proliferation, cell cycle and cell apoptosis of BC and whether the effects are related to ER/GPER signaling pathway. The results demonstrated that shikonin inhibited the cellular proliferation of MCF-7 BC cells via G0/G1 arrest and apoptosis in concentration-dependent manner. The anti-proliferative effect of SK on SK-BR-3 BC cells was associated with apoptosis. Both ERα and GPER were expressed in MCF-7 cells, while ERα were negative and GPER were positive in SK-BR-3 cells. Furthermore, shikonin downregulated the expression of ERα and GPER, and this effect was not affected by the estrogen environment. In addition, shikonin downregulated the EGFR and p-ERK expression in MCF-7 and SK-BR-3, which was also not affected by the estrogen environment. EGFR and p-ERK were still suppressed by co-treatment with the selective GPER against G1 or antagonist G15. In conclusion, these results suggest that shikonin shows anti-tumor effects on MCF-7 and SK-BR-3 cells. The effects seem to be associated with EGFR/p-ERK downregulation via ERα and GPER inhibition.

Objective To observe the effect of Shikonin ( SK) on the proliferation cycle of SiHa cells in human cervical cancer and to explore the mechanism of it. Methods The human cervical cancer SiHa cells were cultured in vitro, and the cell proliferation index and periodic distribution were detected by flow cytometry after different concentrations of SK were acted on the cells. The expression of G protein cou-pled estrogen receptor ( GPER) and Cyclin D1 in SiHa cells were observed by immunohistochemical meth-od. Results After 48 hours of action, the proliferation index of SiHa cells in the E2 group was higher than that in the blank control group. The proliferation indexes of SiHa cells were decreased in 1×10-5, 1×10-6,1×10-7, 1×10-8 mol/L SK groups(P<0.05). Moreover, G0/G1 cells were increased and S phase cells were decreased( P<0.01) . Compared with the blank control group, the expression of GPER and cyclin D1 protein in E2 group were increased. The expression of GPER and cyclin D1 were significantly decreased in a concentration dependent manner of 1×10-5, 1×10-6, 1×10-7, 1×10-8mol/L SK groups(P<0.05). Con-clusion Shikonin can inhibit the proliferation of SiHa cells in human cervical squamous cancer, and the molecular mechanism of its influence on G0/G1 block may be related to GPER and CyclinD1 proteins.

目的:探讨二仙汤含药血清通过AKT途径介导顺铂所致卵巢颗粒细胞凋亡的防护作用及分子机制.方法本实验采用大鼠原代卵巢颗粒细胞体外培养及顺铂造模的方法.通过选取22-24 d的SD大鼠提取原代卵巢颗粒细胞,进行细胞培养并建立顺铂致卵巢颗粒细胞凋亡模型,将成模卵巢颗粒细胞随机分为:模型组、阳性对照组、二仙汤组,加入相应药理血清连续培养48 h后,用TUNEL法检测各组颗粒细胞凋亡情况;蛋白印迹法检测各组akt、p-akt蛋白的表达情况;RT-PCR检测各组akt基因表达情况.结果:通过TUNEL法对各实验组卵巢颗粒细胞凋亡形态学观察发现:与正常组相比,模型组凋亡情况十分明显.阳性对照组与模型组相比凋亡情况改善;二仙汤组与模型组相比较凋亡情况也得到了缓解.蛋白印迹分析显示:akt、p-akt蛋白表达量总体趋势走向、强度、结果相似.顺铂造模后的各实验组中的akt、p-akt蛋白表达量均减少,模型组akt、p-akt蛋白表达量最少;和模型组比较,各治疗组均akt、p-akt蛋白表达量表达增多,其中二仙汤组akt、p-akt蛋白表达量略低于补佳乐组akt、p-akt蛋白表达量.RT-PCR检测akt基因表达情况,与蛋白表达大体一致.结论二仙汤能明显减轻顺铂引起的卵巢颗粒细胞凋亡,其作用机制与激活AKT信号通路有关.

In recent years, the incidence of premature ovarian failure rate has increased.Modern medicine is mainly treated with hormone replacement therapy, the clinical effect is modest and the side effect is big.The traditional Chinese medicine considers the disease is mainly about age is not old, but the Chong-Ren has been weak, Tiangui has dried up, Shen essence and qi is reduced, and premature ovarian failure is induced.Erxian decoction is mainly Tonifying Kidney Yang, improve the level of estrogen in vivo.Play an important role in the clinical treatment of premature ovarian failure.In this paper, hypothalamic gonadotropin, reproductive hormones regulation of the hypothalamic pituitary ovarian axis and granulosa cell level in recent years was reviewed on Erxian decoction and its disassembled prescriptions in the treatment of premature ovarian failure in experimental study.

Professor NIU Jianzhao argues that the pathogenesis of adolescent polycystic ovary syndrome is kidney deficiency, which leads to disorder of qi and blood. While the cold-evil, phlegm and qi-blood stasis are extrinsic factors. The basic treatment of supplementing kidney and nouring essence, satisfying qi and blood and syndrome diffrentiation was established by Professor NIU. She stresses that tonifying kidney yin and yang and regulating qi and blood should be treated simultaneously. Therefore the benefits of supplying yin and yang, regulating qi and blood as well as treating the basic and inessential aspects will be achieved. In addition, according to the adolescent female physiology, Professor NIU found a method to regulate menstruation and promote ovulation: regulate menstruation and arrest pain in menstrual period, nourish kidney yin in follicular phase, regulate qi and blood to promote ovulate, warm kidney yang and relieve qi stagnancy in luteal phase. On this basis, according to the individual differences in patients, change the dosage of herbs, the clinical curative effects will be remarkable.

Objective To observe the influences of Erxian decoction on the proliferation and cell cycle ovarian granulosa cells caused by CDDP. Methods Different doses of Erxian decoction pharmacological serum were injected in ovarian granulosa cells injured by CDDP,normal pharmacological serum was used as blank control,progynova pharmacological serum was used as positive control,and focus on the comparison of changes after joined LY294002 which was PI3K/AKT pathway inhibitor in each group at the same time. Each group of ovarian granulosa cells proliferation was detected by MTT method when pharmacological serum dosing for 24h,48h and 72h. Cell cycle and phase changes in each group were determined by flow cytometry. Results (1) Erxian decoction has proliferation promoting effect on CDDP injury ovarian granulosa cells,and has proliferation effect of ovarian granulosa cells after injured by CDDP and added LY294002 inhibitors. (2)Erxian decoction each dose group and progynova group can rise the proliferation index by enhancing the proportion of S phase granulosa cells and decreasing G0/G1 phase proportion. Conclusions Erxian decoction has significant proliferation promoting effect of CDDP injury ovarian granulosa cells. The possible mechanism on one hand is that it activates or enhances AKT signaling pathways and inhibits cell apoptosis. On the other hand,it perhaps promotes S phase proportion transformation from G0 phase therefore increases the ratio of S phase cells.

Objective:To observe the effect of serum containing Erxian decoction on regulating the expression of bcl-2and bax protein from rats' ovarian granulosa cells induced by cisplatin and to investigate the mechanism of Erxian Decoction serum on apoptosis of ovarian granulosa cells from the perspective of apoptosise.Methods:Thirty SD female rats for 22 to 24 days birth were randomly divided into three groups:the normal saline group,progynova group and the Erxian Decoction group with 10 rats in each group.After 4-day drug administration,rat hearts were taken to prepare drug serum.Ovarian granulosa cells were taken from SD rats and cultured to acquire adherent cells.Then,primary ovarian granulosa cells in rats were induced by cisplatin.After pharmacological action to serum,radio immunity method was used to detect estrogen and progestin and make analysis.Immunohistochemistry was applied to detect the expression of bcl-2 and bax protein in each group and then for image analysis.Results:Test results showed that E2 expression significantly changed in the model group and progynova group (P<0.05) with statistically significant difference,compared with the normal group.Compared with the model group,the expression of E2 increased significantly in the progynova group and the Erxian Decoction group.Compared with the normal group,P expression in the model group was somewhat changed (P<0.05),which showing statistically significant difference.Compared with the model group,P expression in Erxian Decoction group was slightly higher than that of the progynova group,but lower than that of the normal group.Immunohistochemistry results showed that brown granules of bcl-2 and bax protein were expressed in the cytoplasm of ovarian granulosa cells.Compared with the normal group,the expression of bcl-2 in each group under the intervention of cisplatin was decreased,and the model group was the most obvious (P<0.05).Compared with model group,the positive expression of bcl-2 in each drug group was increased,but it was weaker than that in the normal group (P<0.05).The intensity of bax and bcl-2's expression was opposite.Compared with the normal group,Bax expression in each group under the intervention of cisplatin were enhanced and that in the model group especially showed a strong expression (P<0.05).Compared with the model group,the positive expression of the drug groups was decreased,but was higher than that of the normal group (P<0.05) with statistically significant difference.Conclusion:Erxian Decoction can decrease the expression of Bax protein in follicular granulosa cells induced by cisplatin injury and increase the expression of bcl-2 protein.It can inhibit the apoptosis of ovarian granulosa cells and protect the ovarian function by adjusting the expression of bcl-2 and bax.

目的:通过研究二仙汤对卵巢血供和卵泡颗粒细胞分泌AMH、Inhibin的影响,探究其对卵巢储备功能的影响,从而探究二仙汤治疗卵巢早衰的机制.方法:60只SD大鼠随机分为正常组、模型组、阳性对照组和二仙汤组,腹腔注射顺铂建立化疗性损伤大鼠卵巢早衰动物模型.造模后连续4周灌胃药物治疗.HE染色观察卵巢血管的变化;ELISA法测定血清中AMH、Inhibin含量.结果:与正常组比较,模型组卵巢血管减少,血清AMH、Inhibin下降(P＜0.05);二仙汤和西药补佳乐治疗后与模型组比较卵巢血管增多,血清AMH、Inhibin升高(P＜0.05).结论:二仙汤治疗卵巢早衰的机制之一可能是改善卵巢血供,促进颗粒细胞AMH和Inhibin分泌,恢复卵泡正常募集和成熟,减少卵泡闭锁,从而改善卵巢储备功能.

现代医学教育中，中医院校对基础医学实验课教学大都存在着对实验形式和内容的要求越来越高，但实验设备、器材、环境和经费相对落后的矛盾。如果能构建虚拟实验室，恰当地运用现代教育技术的仿真、模拟功能，则可以变抽象为具体，使静态内容动态化，逼真地模拟实验时实验过程中宏观变化中的微观过程，变微观为直观，让学生观察、体验、感受事物发展过程中的微观现象，能够把实验教学中难以描述、难以突破的难点、重点、生动形象地表现出来，激发学生学习的兴趣，调动学生的学习积极性，强化学生对实验学知识的记忆和理解。