己烯雌酚影响新生小鼠睾丸引带相关基因表达的初步研究

目的:通过不同剂量己烯雌酚(DES)孕期染毒,了解其对新生仔鼠睾丸引带中雄激素受体(AR)、雌激素受体α(ERα)、增殖细胞核抗原(PCNA)和肌动蛋白α1(ACTα1)mRNA表达的影响,以期窥探DES影响睾丸引带的作用途径。方法:将发现雌性昆明小鼠阴道栓当天记为孕第0天(GD0),在GD9~GD17分别给予不同剂量DES[0.02、0.1、0.5、10、50μg/(kg·d)]作为实验组,给予等体积二甲基亚砜及生理盐水分别作为溶剂和空白对照组。生后第1天处死雄性仔鼠获取睾丸引带,通过RT-PCR检测AR、ERα、PCNA和ACTα1 mRNA的表达量。结果:随着DES浓度的增加,ERαmRNA的表达逐渐增加,各实验组与溶剂对照组比较差异有统计学意义(RE~2=0.825,P0.05);AR、PCNA和ACTα1 mRNA的表达都呈下降趋势,高剂量组[10、50μg/(kg·d)DES组]与溶剂对照组比较差异有统计学意义(RA~2=0.713,RP~2=0.946,RT2=0.960,P0.01)。结论:正常新生昆明小鼠睾丸引带中存在ERα、AR、PCNA及ACTα1 mRNA的表达。孕期不同剂量DES染毒对新生小鼠睾丸引带细胞受体蛋白(ERα和AR)及功能蛋白(PCNA和ACTα1)mRNA表达的影响不同。外源性雌激素可能通过影响ER和或AR的代谢而影响睾丸引带细胞的增殖和收缩等功能,从而影响睾丸甚至整个雄性生殖系统的正常发育。     

小鼠睾丸引带细胞的培养及己烯雌酚对培养睾丸引带细胞的影响

目的:探讨小鼠睾丸引带细胞体外培养的有效方法,并进行形态学观察。研究经典的外源性雌激素己烯雌酚(DES)体外对小鼠睾丸引带发育的影响。方法:手术放大镜解剖出3日龄雄性昆明小鼠的睾丸引带组织并进行细胞培养。台盼蓝染色检测细胞存活率,HE染色观察细胞形态。传代培养并随机分为空白对照组、溶剂对照组(DMSO组)和实验组(DES 1~4组)共6组。溶剂对照组加入DMSO,实验组分别加入0.01、0.10、1.00以及10.00μg/ml DES。分别于培养12、24、48 h后进行细胞形态学观察,CCK-8法检测睾丸引带细胞的生长情况。结果:培养睾丸引带细胞大部分为成纤维细胞型,有少数的上皮样细胞。原代细胞的存活率为85%~90%。在不同剂量DES作用后的3个不同时间段里,细胞增殖性的检测结果存在时间-剂量效应的显著差异(P<0.01)。结论:睾丸引带细胞可经一定的方法进行体外培养,外源性雌激素对睾丸引带细胞的生长有抑制作用,且呈现一定的时间-剂量效应。对培养睾丸引带细胞影响的研究是外源性雌激素影响生殖系统发育研究的一条有效的睾丸外途径。     

隐睾症睾丸引带异常的临床意义

目的探讨睾丸引带异常的临床意义。方法1994年1月至2003年12月对169例接受手术的隐睾症患者进行术中直接观察,记录隐睾位置、睾丸引带长度和附着点。结果169例212枚隐睾中,腹腔内睾丸42枚(19.8%),腹股沟管睾丸134枚(63.2%),阴囊上部睾丸36枚(17.0%)。腹腔内睾丸引带附着点位于腹股沟管外环周围、阴囊上部和其他位置者分别占73.8%(31)、21.4%(9)和4.8%(2);腹股沟管睾丸引带附着点位于腹股沟管外环周围者64.9%(87),阴囊上部者26.1%(35),阴囊底部者6.7%(9);阴囊上部睾丸引带附着点位于腹股沟管外环周围者25.0%(9),阴囊上部者52.8%(35),阴囊底部者22.2%(8)。腹腔内、腹股沟管和阴囊上部睾丸引带长度分别为(3.1±1.1)、(3.2±0.7)和(2.5±0.4)cm,阴囊上部睾丸引带长度明显短于其他2型隐睾(P<0.01)。结论大多数隐睾有引带异常如迁移停止或未退化或异常附着,这可能是隐睾症激素治疗难以奏效的原因。保留引带睾丸固定术对需要切断精索血管的高位隐睾和精索血供不良的隐睾的血供具有重要价值。     

胰岛素样激素3与睾丸引带发育和睾丸下降

睾丸下降是生殖系统发育过程中的重要阶段,其机制尚未阐明。睾丸引带发育与睾丸下降关系密切。睾丸下降不全常常伴随睾丸引带发育异常,它们除了可引起隐睾和睾丸发育不良等疾病外,还是生殖系统先天畸形以及以后生殖功能、性功能异常和高比例生殖系统肿瘤发生的原因。胰岛素样激素3是近年来倍受关注的影响睾丸引带发育和睾丸下降的重要因素,本文就相关进展进行综述。     

影响睾丸引带发育的内分泌因素研究进展

睾丸引带与睾丸下降关系密切,它的分化发育和多种因素有关,特别是雄激素、降钙素基因相关肽、胰岛素样因子3、苗勒管抑制物质、表皮生长因子以及环境雌激素(EEs)等,其中降钙素基因相关肽、胰岛素样因子3和EEs对生殖系统的影响是目前研究的热点。从影响睾丸引带发育的角度探讨EEs对睾丸及生殖发育的影响很有意义。     

腹横纹切口保留睾丸引带阴囊皮下睾丸固定术治疗隐睾症的研究

为了研究治疗先天性隐睾症的最佳术式，以提高隐睾症的治疗水平，采用腹横纹切口保留睾丸引带明囊皮下睾丸固定术（研究组）治疗隐睾症96例110枚，获得随访者94例108枚，同时采用患倒下腹斜切口肉膜囊睾丸固定术（对照组）治疗隐睾症50例62枚。结果研究组睾丸大小及硬度化94枚，占87．0％，良9枚，占8．3％，差5枚，占4．6％；睾丸位置优93枚，占86．1％，良9枚，占8．3％，差6枚，占5．6％；无睾丸萎缩及回缩，外表美观。对照组睾丸大小及硬度优41枚，占66．1％，良7枚，占11．3％，差14枚，占22．6％；睾丸位置优43枚，占69．4％，良8枚，占12．9％，差11枚，占17．7％。经统计学处理，两组睾丸大小及硬度方面比较有极显著性差异（P＜0．01），睾丸位置比较也有显著性差异（P＜0．05）。认为腹横纹切口保留睾丸引带阴囊皮下睾丸固定术损伤小，外表美观，明显降低了睾丸萎缩及回缩等并发症，符合生理要求，疗效满意。     

己烯雌酚对小鼠睾丸引带中LGR8表达的影响

目的:通过体内实验研究己烯雌酚(DES)对小鼠睾丸引带中胰岛素样因子3受体LGR8的影响,从而探讨外源性雌激素对小鼠睾丸下降的影响。方法:8～10周龄KM孕鼠随机分为正常组、空白对照组和实验组(0.1、1.0、10、100μg/kg DES 4个剂量组),共6组,每组20只。于孕9～17 d每天分别给予实验组妊娠小鼠不同剂量的DES(0.1、1、10、100μg/kg),空白对照组给予等体积DMSO+生理盐水,正常组不给药。应用免疫组化和RT-PCR分别检测胎鼠和幼鼠睾丸引带中LGR8蛋白和mRNA的表达。结果:HE染色可见胎鼠正常组、对照组睾丸引带发育良好,中间的间叶组织和外周的肌源细胞之间分界清楚;实验组可见睾丸引带发育不良,间叶组织和肌源细胞之间无明显分界,组织结构紊乱。幼鼠正常组和实验组睾丸引带发育未见明显不同。LGR8表达于睾丸引带肌源细胞和间叶组织细胞,以肌源细胞表达为主。实验组LGR8阳性表达较正常组弱,胎鼠各实验组和幼鼠DES 1、10、100μg组与同发育阶段的正常组相比均有统计学意义(P<0.05)。DES高剂量(10、100μg)组与同发育阶段的正常组相比LGR8 mRNA表达增加(P<0.05)。各实验组PCR产物测序均未见明显突变。结论:DES可影响小鼠睾丸引带LGR8 mRNA和蛋白的表达,DES可能通过影响INSL3-LGR8信号系统干扰睾丸引带的发育,进而影响睾丸正常下降。     

保留睾丸引带在腹腔镜小儿腹腔型隐睾手术中的应用

目的:探讨保留睾丸引带在腹腔镜小儿腹腔型隐睾手术中的疗效.方法:回顾分析2007年6月至2010年8月为24例腹腔型隐睾患儿行腹腔镜睾丸阴囊肉膜囊固定术的临床资料,术中保留睾丸引带和精索双向血供.结果:24例均成功完成腹腔镜Ⅰ期睾丸下降固定术,无一例中转手术,手术时间45～60 min,平均55 min,无并发症发生....     

结扎睾丸引带静脉对显微精索静脉曲张手术疗效的影响

目的:比较外环下显微精索静脉结扎术提出睾丸结扎引带静脉与不提出睾丸保留引带静脉的疗效及并发症。方法:回顾性总结163例精索静脉曲张患者术前及术后的临床资料,其中提出睾丸40例,不提出睾丸123例。比较不同手术方法在手术时间、术后并发症、复发率,以及术前、术后3个月精液质量的差异。结果:提出睾丸组手术时间(81.1±20.0)min,不提出睾丸组手术时间(62.3±9.6)min,两组间有统计学差异(P=0.041)。两组术后精子存活率、前向运动精子百分率、精子浓度以及1次射精总数均较术前显著改善(P0.05)。其中提出睾丸组Ⅲ度精索静脉曲张32例,不提出睾丸组Ⅲ度精索静脉曲张62例。两组间Ⅲ度曲张患者术前精子存活率、前向运动精子百分率、精子浓度、1次射精精子总数及术后3个月精子存活率、前向运动精子百分率、精子浓度、1次射精精子总数均无统计学差异(P均0.05)。提出睾丸组术后5例患者伴有阴囊水肿,不提出睾丸组术后2例患者伴有伤口感染。两组患者均无睾丸萎缩及精索静脉曲张复发。结论:外环下显微精索静脉结扎术提出睾丸结扎引带静脉与不提出睾丸保留引带静脉相比较,并没有更好的减少复发率和提高精液参数,反而更加延长手术时间以及引起阴囊水肿等并发症的发生。     

邻苯二甲酸二(2-乙基)己酯对小鼠胎鼠睾丸及睾丸引带的影响

目的:研究邻苯二甲酸二(2-乙基)己酯(DEHP)对小鼠胎鼠睾丸与睾丸引带形态结构及功能的影响,探讨隐睾发生的可能机制。方法:30只健康KM孕鼠随机均分为3组:空白对照组、玉米油对照组、DEHP组。DEHP组以剂量500mg/(kg.d)的DEHP灌胃作用于怀孕12～19d(GD12～GD19)的KM母鼠,观察DEHP对每胎胎鼠数、雌雄性胎鼠比例、雄性胎鼠体重、睾丸重量、睾丸引带形态结构、位置、睾丸到膀胱颈之间的相对距离(TBD)、颅侧悬韧带残留情况、引带内雄激素受体(AR)、雌激素受体(ER)、肌动蛋白、增殖细胞核抗原(PCNA)表达水平的影响。结果:DEHP对每胎胎鼠数、雌雄性胎鼠比例、雄性胎鼠体重无明显影响;DEHP可影响胎鼠睾丸重量及TBD;DEHP组睾丸均有一定程度的下降不全,睾丸引带形态结构无明显差异;光镜下见DEHP组睾丸生精小管、支持细胞存在明显的发育障碍,睾丸Leydig细胞明显增生;DEHP组睾丸引带AR阳性表达率降低(P<0.01)。结论:DEHP可通过抗雄激素效应导致睾丸引带功能障碍,使睾丸下降发生异常而诱导小鼠隐睾;同时造成睾丸Ser-toli细胞、睾丸Leydig细胞和生精细胞的发育障碍和功能改变。     

雷公藤多甙对小鼠生育的影响及肉苁蓉干预作用的研究

目的:研究使用不同时间和剂量的雷公藤多甙(GTW)对昆明小鼠怀孕率的影响及肉苁蓉干预和停用GTW后的自愈倾向。方法:将90只成年雄性昆明鼠随机分为9组(每组10只)。对照组给予1%羧甲基纤维素(CMC)溶液;GTW各组分别给予45.0、30.0、22.5、15.0、7.5mg/(kg·d)GTW混悬液;肉苁蓉各组分别给予30.0、7.5mg/(kg·d)GTW混悬液;停药组给予30.0mg/(kg·d)GTW混悬液。以上各组先灌胃20d,20d后与成年雌鼠1∶2合笼7d。第28d起GTW各组按原剂量继续给药;肉苁蓉各组给原剂量GTW混悬液的同时分别给10.0、2.5g/(kg·d)肉苁蓉煎液;停药组停GTW,改为1%CMC溶液。20d后再次合笼7d。每次合笼后观察雌鼠怀孕率。结果:随着GTW给药时间的延长和剂量的增加,雌鼠怀孕率呈下降趋势。GTW45.0mg/(kg·d)组给药40d后怀孕率降为0%(P<0.01)。肉苁蓉10.0g/(kg·d)组怀孕率由20d时的25%上升到40d时的55%。停药组怀孕率由25%上升到56%,与对照组比较,差异无显著性(P>0.05)。结论:GTW对雄性小鼠的生殖毒性呈时间和剂量依赖性。肉苁蓉可以改善GTW对雄鼠生殖系统的抑制作用,提高雌鼠的怀孕率。停用GTW20d,怀孕率可以基本恢复正常。     

Morphometric Measurements of the Cranium in Patients with Chiari Type I Malformation and Comparison with the Normal Population

Summary. Summary.   Background: To determine the degree of development of the posterior fossa and signs of occipital dysplasia in patients with Chiari type I malformation by morphometric measurements.   Methods: In 22 patients with Chiari type I malformation, distance, surface area and angle values reflecting the degree of development of the posterior fossa were measured and compared with the measurements of 21 normal subjects.   Findings: In patients with Chiari type I malformation, the depth of the posterior fossa, the length of the clivus reflecting development of the basi-occiput and Klaus' index were significantly shorter than in normal subjects (p<0.001, p=0.007, and p<0.001, respectively). The ratios of the depth of the posterior fossa to the height of the supratentorial region and Twining's line reflecting anteroposterior distance of the posterior fossa were also significantly smaller in the Chiari group (p<0.001 for both). In sagittal section, the surface area of the bony part of the posterior fossa and the ratio of this area to the area of the supratentorial region were significantly smaller than in normal subjects (p=0.038 and p=0.002, respectively). The angle measurements of the cranial base (basal angle, Boogard's angle and nasion-basion-opisthion angle) showed that there was an evident tendency for platibasia in the Chiari group (p=0.04, p=0.004, p<0.001, respectively). In addition, it was shown by measuring tentorium-twining's line angle that the tentorium was steeper in the Chiari group than normal subjects.   Interpretation: These results support the opinion, which claims the existence of underdevelopment of the occipital bone and posterior fossa in patients with Chiari type I malformation.     

急性热应激对性成熟雄性小鼠睾丸、附睾、输精管中热休克蛋白70表达的影响

目的:探讨急性热应激对性成熟雄性小鼠睾丸、附睾、输精管中热休克蛋白70(heat shock protein 70,HSP70)表达的影响。方法:将32只8周龄雄性小白鼠随机均分为4组,饲养7d后,进行热应激处理,温度控制在(39±0.5)℃,时间分别为0.5、1和3h。应激后立即采血,分离血清测定谷草转氨酶(GOT)含量。一侧附睾制备精子悬液,用于计算精子密度和顶体畸形率;另一侧附睾、睾丸、输精管用于免疫组化研究。结果:应激后,小鼠体重、睾丸系数、顶体畸形率变化不显著(P>0.05),附睾系数和精子密度有不同程度的下降,GOT含量急剧升高(P<0.01)。随着应激时间的延长,小鼠精子密度呈递减趋势,顶体畸形率呈上升趋势。应激时间最短的0.5h组小鼠体重、睾丸系数、附睾系数的降幅反而最大。免疫组化法观察发现,HSP70在性成熟小鼠睾丸、附睾、输精管中均有表达。正常状态下,HSP70在睾丸组织间质细胞中少量表达,应激后分布于间质细胞核,此外在精母细胞核与精子细胞核中也有大量分布;附睾中HSP70主要分布于主细胞质,基细胞和亮细胞中没有表达,应激后附睾体的纤毛细胞中也发现大量棕色颗粒;输精管中HSP70主要定位在基细胞质,主细胞中不表达。随着应激时间的延长,HSP70在睾丸、附睾中的表达量明显升高,而在输精管中的增幅不明显。结论:急性热应激对性成熟雄性小鼠的生殖系统造成了损伤;HSP70在睾丸、附睾、输精管中的表达与定位具有区域特异性和细胞特异性,提示其可能参与精子的发生与成熟;HSP70在应激状态下表达量大幅上升的作用可能在于保护细胞免受高热损伤。     

Stage-Dependent Enzymatic Activities in Spermatogenesis of Mice with the Standard Karyotype and of Chromosomal Variants with Impaired Fertility

The enzymatic activities of thiamine pyrophosphatase (TPPase), acid phosphatases (ACPases), alkaline phosphatases (APases) and steroid-3 beta-ol dehydrogenase (beta-HSDH) in different germ cells and somatic cells in the testis of three cytogenetically determined states of fertility (i.e. normal, impaired fertility and sterility) were studied histochemically in the mouse. The TPPase, ACPases and APases activities showed a characteristic stage dependent pattern when the activities were related to the typical twelve stages of the seminiferous epithelium in the mouse, according to Oakberg (1956). Comparing the enzymatic patterns of the activities in the normal spermatogenic process versus the impaired and sterile conditions, the following conclusions can be drawn: even in impaired and sterile conditions the enzymatic activity patterns retain their characteristic stage dependence; the pattern of beta-HSDH and ACPases is not altered in the impaired and sterile conditions; TPPase and APases patterns are modified in impaired and sterile mice. It is concluded that the kinetics of the enzyme activities can serve as a useful marker for characterizing pathological spermatogenic processes.     

Chiari malformations in adults: A single center surgical experience with special emphasis on the kinetics of clinical improvement

BackgroundThe Chiari malformation type I (CM-I) is the most commonly found type in adults. The efforts to further improve the treatment offered for these malformations are hampered by the existence of controversial methods and the absence of a uniform scoring system to evaluate clinical outcomes.ObjectivesThe goal of our study is to analyze the clinical and radiological data concerning patients operated for CM and to expose surgical techniques.Patients and treatmentThis is a retrospective study concerning patients of more than 16 years of age, operated on (from 2000 to 2016) in our institution. These patients underwent bony decompression of the cervico-occipital junction, with a duraplasty enlargement. Clinical and radiological follow-up was assessed sequentially.ResultsThe mean age of patients included in this study was 39. Headaches (n = 19) and sensory disturbances (n = 17) were the most common presenting complaints. Furthermore, syringomyelia symptoms were present in 34.5% of the cases (n = 10). Twenty-three patients displayed a Chiari malformation of type I (79.3%), and six patients were classified as Chiari malformation type I.5 (20.7%). A syringomyelia was present in 58.6% of the cases (n = 17). The postoperative complications that were encountered were: one case of pseudo-meningocele, two cases of cerebrospinal fluid leakage, two cases of meningitis, and one case of delayed wound healing. The mean follow-up period was 18 months, which showed beneficial outcomes in 82.8% of the cases (20.7% cured, and 62.1% improved) and an unfavorable outcome in 17.2% of the cases (13.8% stable patients and 3.4% worsened outcomes). Syringomyelia symptoms were improved in 60% of the patients. Among the patients who presented without a syrinx, 82.3% had good outcomes; and those who presented with a syrinx, 83.4% had good outcomes. Symptoms improved for 69% of patients within 3 months.ConclusionAn optimal craniocervical osteo-dural decompression plus duraplasty offered early and sustainable good clinical results in symptomatic CM-I and CM-I.5 patients.     

A Long-term Study of the Epididymal- and Fertility-Suppressing Effects of Cyproterone Acetate in the Mouse

The influence of cyproterone acetate (CA) (0.1 mg/mouse/day) on the epididymis and male fertility was analyzed in a long-term study in the Swiss albino cc strain. CA inhibits epididymal function in all segments as measured by organ weights, protein content and beta-glucuronidase activity. This antiandrogen also suppresses male fertility. It is important to note that the low dose of CA, used in this study, does not affect spermatogenesis. The effects of CA on epididymis and fertility are reversible and the magnitude of recovery or the period (in days) required for a significant recovery after the discontinuation of CA were seemingly directly proportional to the duration of antiandrogen treatment.     

Syringomyelia Associated with Type I Chiari Malformation A 21-Year Retrospective Study on 75 Cases Treated by Foramen Magnum Decompression with a Special Emphasis on the Value of Tonsils Resection

Summary  The purpose of the present study is to evaluate retrospectively the effects of several intra-operative manipulations on the results of foramen magnum decompression (FMD) in patients having syringomyelia associated with type I Chiari malformation. Seventy-five patients having syringomyelia associated with Chiari I malformation were operated on between 1975 and 1996. This population was grouped into 4 subgroups according to the surgical protocol: group I=42 patients with FMD alone; group II=16 patients with FMD and third ventricle shunting; group III=9 patients with FMD and syringosubarachnoid shunting (SSS); group IV=8 patients with FMD and cerebellar tonsils resection. Pre- and postsurgical magnetic resonance imaging (MRI) studies were analyzed (and compared). Nine patients were lost to follow-up. The results were evaluated in the 66 remaining patients (mean follow-up: 52 months), using the Bidzinski's outcome scale (ref). Two patients (3%) died postoperatively, 31 (47%) had very good results (after additional surgery in 7), 16 (24,2%) had good results (after additional surgery in 7) and 17 (25,7%) had poor results despite further surgery in 9. A total of 27 reoperations were undertaken after primary FMD in 23 patients (35%). Thirty-nine patients (59%) had both pre- and postsurgical MRI evaluation. In 28 (72%) the syrinx had markedly decreased whereas it had remained stable in 11 (28%). Clinical results were not significantly different between the patients of groups I, II and III. Very good or good results were obtained in 24 patients (64,8%) of group I (after additional surgery in 10), in 8 (61,5%) of group II (after additional surgery in 1) and in 7 (87,5%) of group III (after additional surgery in 3). Results in group IV were as follows: 7 patients (87%) had very good results and one had a good result. With a mean follow-up of 28 months, no patient required additional surgery. Postsurgical MRI syrinx reduction was observed in all 8 patients either in the early postoperative course or on delayed follow-up. It is suggested that tonsils resection might enhance the results of FMD in individuals having Chiari I-related syringomyelia.     

肝硬化,门静脉狭窄大鼠胃粘膜屏障功能异常的研究

为观察门静脉高压时胃粘膜的屏障功能，研究设计肝硬化和肝外门静脉狭窄两组门静脉高压动物模型，观察其胃粘膜屏障功能的变化并与正常对照组对比研究。结果显示：门静脉高压大鼠内脏血流量明显增加，而胃粘膜实际处于局部缺血状态；胃壁结合粘液量（ＧＰ）和胃粘膜ＰＧＥ２含量较对照组显著降低，尤以肝硬化门静脉高压大鼠为甚；胃基础泌酸量（ＢＡＳ）三组间无差异，而门静脉高压大鼠Ｈ＋返流量（Ｈ＋ＢＤ）明显高于对照组，以肝硬化大鼠最为显著。结果表明：门静脉高压性胃病（ＰＨＧ）的发生与胃粘膜屏障功能严重削弱有关；而非“高酸”所致；肝功能受损参与胃粘膜病变的发生。