未成熟卵体外培养发育潜能探究

目的体外受精-胚胎移植(In vitro fertilization and embryo transplantation,IVF-ET)周期所获卵子均未成熟时,其体外培养发育潜能探究。方法 25例IVF-ET周期所获卵子均未成熟(未成熟卵149枚),通过实验室体外过夜培养所有未成熟卵子(M1期,142枚;GV期,7枚),培养成熟卵子(M11期,98枚)行体外单精子胞内注射(Intracytoplasmic sperm injection,ICSI)后进行72h体外培养;回顾性分类统计IVF-ET周期中IVF、ICSI和Half ICSI的正常受精率、正常卵裂率和优胚率,比较它们间的差异。结果 IVF周期与Half ICSI周期中的IVF部分在受精率、正常卵裂率和优胚率均无统计学差异;ICSI周期与Half ICSI周期中的ICSI部分在受精率、正常卵裂率和优胚率均无统计学差异;所获卵子均未成熟周期与ICSI周期在受精率和优胚率有统计学差异,在正常卵裂率无统计学差异;所获卵子均未成熟周期与HalfICSI周期中的ICSI部分在受精率和优胚率有统计学差异,在正常卵裂率无统计学差异。结论所获卵子均未成熟周期的未成熟卵子经体外培养能部分成熟,并具有继续发育潜能,但其受精率和优胚率均明显低于体内成熟卵子的相应指标;且他们最终临床结局较差。     

卵母细胞第一极体形态与女性年龄及ICSI结局的相关性研究

目的探讨卵母细胞第一极体形态与女性年龄及ICSI结局的相关性。方法回顾性分析了2008年3月至2009年11月在我院行ICSI治疗的146个周期的临床资料,按MⅡ期卵母细胞第一极体形态分为完整极体组（1、2级极体）和异常极体组（3、4级极体）,比较不同形态第一极体与年龄及ICSI结局的相关性。结果 35岁以上女性1级和2级极体的卵母细胞比例显著低于小于35岁患者（P〈0.01）;极体完整组的受精率、卵裂率、早裂率、优胚率均显著高于极体异常组（P〈0.01）;移植完整极体胚胎组的着床率和临床妊娠率显著高于移植混合胚胎（含完整极体和异常极体胚胎）组（P〈0.01）。结论卵母细胞第一极体形态与年龄间存在一定的相关性,第一极体形态可作为选择移植胚胎的指标之一。     

人卵母细胞胞浆形态与卵胞浆内单精子显微注射受精率与胚胎质量的关系

目的了解人卵母细胞胞浆形态与卵胞浆内单精子显微注射(ICSI)后正常受精率、卵裂率和优质胚胎率的关系;方法选择2014年2月至2014年8月行ICSI治疗的110个周期中1017枚人卵母细胞。按卵母细胞内是否含有特殊形态结构分为:无特殊结构卵子(A)和含有折光体(B1)、滑面内质网小囊聚集体(SERC)(B2)、空泡(B3)和胞浆颗粒异常(B4)。通过卵子准备、精子准备和ICSI操作过程,了解卵母细胞正常受精、卵裂和培养后优质胚胎率;结果 A组与B3组正常受精率分别为75.98%和42.31%,两组比较有显著性差异(χ~2=29.469,P0.05)。结论不含有空泡的卵子通过ICSI授精,可以得到较高的受精率,卵子内其他一些异常结构对受精及胚胎发育的影响还需要大样本的研究。     

黄体中期长方案及拮抗剂方案对IVF患者卵母细胞质量的影响

目的比较两种控制性超排卵方案对卵母细胞质量的影响。方法回顾性分析2012年1月至2014年7月在北京妇产医院生殖医学科接受黄体中期长方案317例(326个周期)及拮抗剂方案218例(220个周期)治疗的患者,对其卵母细胞质量进行评级,比较两组卵母细胞质量、成熟率、受精率、卵裂率及优质胚胎率等。结果两组患者年龄、不孕年限、BMI、基础FSH、LH、E2水平、Gn总量均无统计学差异(P0.05)。黄体中期长方案组获卵数、成熟卵子数,3级卵子数显著高于拮抗剂组(P0.05);受精数、正常受精数、卵裂数、优胚数均显著高于拮抗剂组(P0.05)。两组的卵母细胞成熟率及各级成熟率、正常受精率、卵裂率、优胚率差异无统计学意义(P0.05)。结论黄体中期长方案组的获卵数、优卵数及优胚数均高于拮抗剂方案组,但两组间成熟卵子率、卵裂率、及优胚率均无差异,两种方案对患者卵母细胞质量影响差异不明显。     

第二极体排出时间早晚与受精率、胚胎质量的关系

目的探讨第二极体(PB2)排出时间早晚与受精率、胚胎质量的关系。方法选择2012年11月~2013年10月在广东省第二人民医院生殖医学科行常规体外受精周期的患者,于授精后的4h观察PB2排出情况,4h内排出PB2为A组,4h内未排出PB2为B组。比较两组间的受精率、卵裂率、可利用胚胎率和优胚率。结果 1A组的受精率、两原核(two pronuclear,2PN)率显著高于B组(P0.05);但两组间的异常受精率、多精受精率和卵裂率无明显差异。2A组的可利用胚胎率及优质胚胎率显著高于B组(P0.05)。结论受精4h内排出第二极体的卵子的总受精率、正常受精率、可利用胚胎率及优质胚胎率显著高于4h内未排出第二极体的卵子,第二极体排出有助于预测体外受精结局。第二极体排出时间与胚胎质量存在一定关系,有助于预测胚胎的发育潜能。     

短时受精不同脱颗粒细胞方式对体外受精结局的影响

目的探讨体外受精-胚胎移植（IVF-ET）治疗中几种短时受精脱颗粒细胞方式对胚胎质量及临床结局的影响。方法将261个IVF-ET周期根据脱颗粒细胞时间分为：短时受精后18小时脱颗粒细胞组（A组）94个周期、短时受精后5h脱颗粒细胞组（B组）95个周期和短时受精5h后部分（1/3-1/2）脱颗粒细胞组（C组）72个周期。分别比较三组的受精率、多精受精率、卵裂率、优胚率、胚胎利用率、临床妊娠率、种植率和流产率。结果三组的受精率、正常受精率、多精受精率、卵裂率、临床妊娠率和流产率差异无统计学意义（P〉0.05）,A组的优胚率、胚胎利用率和胚胎种植率显著高于B组（P〈0.05）。结论短时受精后即刻剥除部分卵子颗粒细胞观察第二极体,在确保受精的情况下剩余卵母细胞保留自体卵周颗粒细胞共培养,可以提高常规IVF的胚胎质量,从而提高常规IVF总的临床妊娠率和胚胎种植率。     

单卵母细胞对应的卵丘颗粒细胞中生长分化因子-9和骨形成蛋白-15的表达水平与胚胎发育潜能的研究

目的评估单卵母细胞对应的卵丘颗粒细胞中GDF9和BMP15的m RNA表达水平对卵母细胞发育潜能的影响。方法采用长方案治疗25例ICSI助孕的女性,取卵当天收集180份单卵母细胞对应的卵丘颗粒细胞,随后提取卵丘颗粒细胞的m RNA,进行实时荧光定量分析,对GDF9和BMP15 m RNA基因表达水平采用独立样本t检验分析。结果常规长方案CCs 180份,因卵子与收集的卵丘颗粒细胞是一一对应的,根据卵子受精情况将CCs分为正常受精组CCs 129份,异常受精组CCs 51份;正常受精组根据第3日胚胎卵裂情况分为优质卵裂胚形成组CCs 58份,非优质卵裂胚形成组CCs71份;非优质卵裂胚继续培养,根据第5日形成囊胚情况,分为优质囊胚形成组CCs 25份,非优质囊胚形成组CCs 46份。GDF9和BMP15 m RNA基因表达水平与正常受精率,优质胚胎率显著相关。正常受精组GDF9和BMP15 m RNA基因表达水平显著高于非正常受精组(P0.01),优质胚胎组显著高于非优质胚胎组(P0.01)。结论单卵母细胞对应的卵丘颗粒细胞中GDF9和BMP15 m RNA基因表达水平与正常受精率,优质胚胎率和优质囊胚形成率呈显著正相关,提示其可以作为预测卵母细胞的发育潜能的指标。     

分析卵母细胞MII率对ICSI受精及胚胎发育的预测价值

目的分析卵母细胞MⅡ率与ICSI后受精及胚胎发育的关系。方法选取2015年4月至2017年7月在本中心接受Gn RH激动剂(Gn RH-a)促排卵的ICSI治疗患者792例,获卵数≥5枚且≤30枚。首先分析标准方案人群609例,根据卵母细胞MⅡ率的不同将患者分为A组:MⅡ率90%(N=220)。B组:90%≥MⅡ率80%(N=179)。C组:80%≥MⅡ率70%(N=124)。D组:70%≥MⅡ率50%(N=66)。E组:50%≥MⅡ率10%(N=20)。以A组为对照,分析BCDE组随着MⅡ率的逐渐降低ICSI后的受精率、2PN受精率、卵裂率、2PN卵裂率、优胚率、2PN优胚率的变化趋势。再根据上述分析结果,将全部入选人群分为高MⅡ率组和低MⅡ率组,分析比较ICSI后的受精率、2PN受精率、卵裂率、2PN卵裂率、优胚率、2PN优胚率的差异。结果标准方案人群,5组患者基本资料无差异的前提下,随着MⅡ率逐渐降低(从A组到E组),受精率、2PN受精率、卵裂率、2PN卵裂率、优胚率、2PN优胚率均呈现逐步降低的趋势。其中,C组、D组的受精率显著低于对照组A组(P0.013)。2PN受精率降低幅度不显著(P0.013),异常受精率没有明显波动。卵裂率以及2PN卵裂率呈现相类似的趋势,D组显著低于A组(P0.013)。优胚率以及2PN优胚率逐渐降低,D组显著低于A组(P0.013)。在Gn RH-a促排患者中,根据前述分析结果,以70%的MⅡ率为分界,受精率、2PN受精率、卵裂率、2PN卵裂率、优胚率、2PN优胚率呈显著差异(P0.05)。结论MⅡ率与ICSI后的受精及胚胎发育结局密切相关,MⅡ率降低,受精率、卵裂率、优胚率均呈现下降趋势。尤其是MⅡ率低于70%,不同方案患者受精及胚胎发育均显著降低。MⅡ率可以作为预测ICSI后受精及胚胎发育结局的一个有效指标。     

分析短时受精及早补救ICSI的应用价值

目的分析短时受精后观察第二极体及受精失败后早补救ICSI在体外受精-胚胎移植中的应用价值。方法回顾分析本中心2013年1月—2015年5月第一次行常规IVF-ET治疗周期的患者资料,根据受精后3h是否剥除颗粒细胞观察第二极体分为第二极体观察组(605例),非第二极体观察组(217例),分析两组的数据;并对比受精失败组与同期ICSI周期的数据进行分析。结果 1)短时受精后第二极体观察对正常受精率、多精受精率、优质胚胎率及临床妊娠率无明显影响(P0.05),2)受精失败组的基础LH水平、成熟卵率明显低于正常受精组(P0.05),3)对比低受精组、完全受精失败组与常规ICSI周期,分析后发现完全受精失败组的成熟卵率及优质胚胎率均明显低于低受精失败组和常规ICSI组(P0.05),正常受精率和临床妊娠率无统计学差异。结论短时受精后剥除颗粒细胞观察第二极体联合补救ICSI能有效的避免受精失败并减少ICSI率,患者的基础LH水平及获卵的成熟度可能会影响受精及胚胎的发育。     

ICSI操作中拆卵时间对胚胎质量及妊娠结局的影响

目的探讨人类卵胞浆内单精子注射（ICSI）技术中,取卵后去除卵周颗粒细胞（拆卵）的适合时间。方法选择09年6月至10年3月男方精液正常、因输卵管因素进入周期的ICSI病例共190对夫妇作为研究对象。随机分为2组,分别在取卵后1～2h（G1）和4～6h（G2）拆卵行ICSI。回顾性分析2组卵子成熟率、受精率、卵裂率、囊胚形成率及妊娠率。结果两组在获卵数、卵子成熟率、受精率及卵裂率方面无统计学差异;妊娠率G2组高于G1组,但无统计学意义（P〉0.5）;两组囊胚形成率差异显著（P〈0.005）,G2组高于G1组。结论卵周颗粒细胞的存在对卵子体外成熟尤其是细胞质的成熟是必须的,取卵后培养4～6h拆卵行ICSI胚胎质量好,囊胚形成率高,有更好的妊娠结局。     

Pain and Effusion and Quadriceps Activation and Strength

Context:

Quadriceps dysfunction is a common consequence of knee joint injury and disease, yet its causes remain elusive.

Objective:

To determine the effects of pain on quadriceps strength and activation and to learn if simultaneous pain and knee joint effusion affect the magnitude of quadriceps dysfunction.

Design:

Crossover study.

Setting:

University research laboratory.

Patients or Other Participants:

Fourteen (8 men, 6 women; age = 23.6 ± 4.8 years, height = 170.3 ± 9.16 cm, mass = 72.9 ± 11.84 kg) healthy volunteers.

Intervention(s):

All participants were tested under 4 randomized conditions: normal knee, effused knee, painful knee, and effused and painful knee.

Main Outcome Measure(s):

Quadriceps strength (Nm/kg) and activation (central activation ratio) were assessed after each condition was induced.

Results:

Quadriceps strength and activation were highest under the normal knee condition and differed from the 3 experimental knee conditions (P < .05). No differences were noted among the 3 experimental knee conditions for either variable (P > .05).

Conclusions:

Both pain and effusion led to quadriceps dysfunction, but the interaction of the 2 stimuli did not increase the magnitude of the strength or activation deficits. Therefore, pain and effusion can be considered equally potent in eliciting quadriceps inhibition. Given that pain and effusion accompany numerous knee conditions, the prevalence of quadriceps dysfunction is likely high.Key Words: arthrogenic muscle inhibition, central activation failure, voluntary activation, muscles

Key Points

• Knee pain and effusion resulted in arthrogenic muscle inhibition and weakness of the quadriceps.
• The simultaneous presence of pain and effusion did not increase the magnitude of quadriceps dysfunction.
• To reduce arthrogenic muscle inhibition and improve muscle strength, clinicians should employ interventions that target removing both pain and effusion.

Quadriceps weakness is a common consequence of traumatic knee joint injury^1,2 and chronic degenerative knee joint conditions.^3,4 Arthrogenic muscle inhibition (AMI), a neurologic decline in muscle activation, results in quadriceps weakness and hinders rehabilitation by preventing gains in strength.⁵ The inability to reverse AMI and restore muscle function can lead to decreased physical abilities,⁶ biomechanical deficits,⁷ and possibly reinjury.⁵ Furthermore, researchers^8,9 have suggested that quadriceps weakness resulting from AMI may place patients at risk for developing osteoarthritis in the knee. In light of the substantial influence of quadriceps AMI on these clinically relevant outcomes, we need to improve our understanding of the factors that contribute to this neurologic decline in muscle activity so efforts to target and reverse it can be implemented and gains in strength can be achieved more easily.Joint injury and disease are accompanied by numerous sequelae (ie, pain, swelling, tissue damage, inflammation), so ascertaining which one ultimately leads to neurologic muscle dysfunction is difficult. Whereas a joint effusion can result in AMI,^10–12 the effects of pain are less understood despite many clinicians attributing AMI to pain. Using techniques that introduce knee pain without accompanying injury may provide insights into the role of pain in eliciting AMI.The degree of knee joint damage may play a role in the quantity of AMI that manifests. Hurley et al^13,14 demonstrated that quadriceps AMI, measured using an interpolated-twitch technique, was greater in patients with extensive traumatic knee injury (eg, fractured tibial plateau, ruptured medial collateral ligament, and medial meniscectomy) than patients with isolated joint trauma (ie, isolated anterior cruciate ligament [ACL] rupture). Similarly, patients with more knee joint symptoms (ie, greater number of symptoms and increased severity of symptoms) may present with greater magnitudes of quadriceps inhibition. Recently, investigators¹⁵ have suggested that patients with more pain display less quadriceps strength, supporting this tenet. Given that effusion and pain often present simultaneously with joint injuries and diseases, such as ACL injury and osteoarthritis, examining both the isolated and cumulative effects of these sequelae appears warranted to determine if they influence the magnitude of muscle inhibition.Experimental joint-effusion and pain models are safe and effective experimental methods that allow for the isolated examination of their effects on muscle function. The effusion model, whereby sterile saline is injected directly into the knee joint capsule,⁷ produces a clinically relevant magnitude of the joint effusion that may be present with traumatic injury. Effusion is thought to activate group II afferents responding to stretch or pressure,^16–18 which in turn may facilitate group Ib interneurons and result in quadriceps AMI.⁵ The pain model involves injecting hypertonic saline into the infrapatellar fat pad to produce anteromedial knee pain similar to that described in patients with patellofemoral pain syndrome.¹⁹ Pain is considered to initiate AMI through activation of group III and IV afferents that act as nocioceptors to signal damage or potential damage to joint structures.^16–18 The firing of these afferents then may lead to facilitation of group Ib interneurons, the flexion reflex, or the gamma loop, ultimately resulting in quadriceps inhibition.²⁰ Thus, these models allow us to create symptoms that are associated with knee injury and have the added benefit of providing a way to examine their effects in isolation.Therefore, the purpose of our study was to determine the effects of pain on quadriceps strength and activation and to learn if simultaneous pain and knee joint effusion would affect the magnitude of quadriceps dysfunction. We hypothesized that pain alone would result in quadriceps inhibition and that the magnitude of inhibition would be greater when effusion and pain were present simultaneously.     

即早基因c-fos与脑血管病及学习记忆

即早基因c-fos是广泛存在于原核细胞和真核细胞的高度保守基因.在正常情况下,c-fos基因参与细胞生长、分化、信息传递、学习和记忆等生理过程,而在病理情况下c-fos基因表达及调控变化与多种疾病的发生和发展有关.C-fos在中枢神经系统的某些部位可有基础水平的表达,但表达很低,当受到如脑缺血、脑出血、痫性发作、应激等刺激后,其在数十分钟内做出反应,在对外界刺激-转录耦联的信忠传递过程中起着核内第三信使的重要作用.     

Opportunities and challenges in the prevention and control of cancer and other chronic diseases: children's diet and nutrition and weight and physical activity

OBJECTIVE: The purpose of this article is to review the role of behavioral research in disease prevention and control, with a particular emphasis on lifestyle- and behavior-related cancer and chronic disease risk factors--specifically, relationships among diet and nutrition and weight and physical activity with adult cancer, and tracking developmental origins of these health-promoting and health-compromising behaviors from childhood into adulthood. METHOD: After reviewing the background of the field of cancer prevention and control and establishing plausibility for the role of child health behavior in adult cancer risk, studies selected from the pediatric published literature are reviewed. Articles were retrieved, selected, and summarized to illustrate that results from separate but related fields of study are combinable to yield insights into the prevention and control of cancer and other chronic diseases in adulthood through the conduct of nonintervention and intervention research with children in clinical, public health, and other contexts. RESULTS: As illustrated by the evidence presented in this review, there are numerous reasons (biological, psychological, and social), opportunities (school and community, health care, and family settings), and approaches (nonintervention and intervention) to understand and impact behavior change in children's diet and nutrition and weight and physical activity. CONCLUSIONS: Further development and evaluation of behavioral science intervention protocols conducted with children are necessary to understand the efficacy of these approaches and their public health impact on proximal and distal cancer, cancer-related, and chronic disease outcomes before diffusion. It is clear that more attention should be paid to early life and early developmental phases in cancer prevention.