阻断部分臂丛神经下干分支对健侧颈7移位下干后神经再生的影响

Objective To explore the changes of the nerve fibers from median and ulnar nerves after cutting the branches of lower trunk which was repaired by the contralateral C7.Methods Forty female SD rats were divided into A, B, C and D groups randomly.In group A,the contralateral C7 root was transferred to lower trunk directly, and the posterior division of lower trunk, medial anterior thoracic nerve and the medial antebrachial cutaneous nerve were severed at the beginning of them;In group B, the contralateral C7 root was trarsferred to lower trunk directly, and the posterior division of lower trunk, medial anterior thoracic nerve and the medial antebrachial cutaneous nerve were severed at the point which was 1 cm away from the beginning of above branches;In group C, the contralateral C7 root was transferred to lower trunk directly, and the posterior division of lower trunk was severed at the beginning of it;In group D, control group.After the operation, myelinated fiber count, nerve fiber density, the percentage of the number of nerve fiber from branch accounting for that from lower trunk, nerve fiber diameter,the percentage of nerve fibers with different diameters and N Ratio were carried out to evaluate the outcome of each group.Results Myelinated fiber count, nerve fiber density, the percentage of the number of nerve fiber from branch accounting for that from lower trunk, nerve fiber diameter,the percentage of nerve fibers with different diameters and N Ratio in ulnar and median nerve, there were no difference between group A, group B and group C ( P ＞ 0.05).Conclusion After the medial anterior thoracic nerve and medial antebrachial cutaneous nerve, repaired by the contralateral C7, were severed at the beginning and at the point which was 1 cm away from the beginning of above branches, the changes of the quantity and quality of the nerve fibers from median and ulnar nerves were not significant.     

阻断部分臂丛神经下干分支对健侧颈7移位下干后神经再生的影响

Objective To explore the changes of the nerve fibers from median and ulnar nerves after cutting the branches of lower trunk which was repaired by the contralateral C7.Methods Forty female SD rats were divided into A, B, C and D groups randomly.In group A,the contralateral C7 root was transferred to lower trunk directly, and the posterior division of lower trunk, medial anterior thoracic nerve and the medial antebrachial cutaneous nerve were severed at the beginning of them;In group B, the contralateral C7 root was trarsferred to lower trunk directly, and the posterior division of lower trunk, medial anterior thoracic nerve and the medial antebrachial cutaneous nerve were severed at the point which was 1 cm away from the beginning of above branches;In group C, the contralateral C7 root was transferred to lower trunk directly, and the posterior division of lower trunk was severed at the beginning of it;In group D, control group.After the operation, myelinated fiber count, nerve fiber density, the percentage of the number of nerve fiber from branch accounting for that from lower trunk, nerve fiber diameter,the percentage of nerve fibers with different diameters and N Ratio were carried out to evaluate the outcome of each group.Results Myelinated fiber count, nerve fiber density, the percentage of the number of nerve fiber from branch accounting for that from lower trunk, nerve fiber diameter,the percentage of nerve fibers with different diameters and N Ratio in ulnar and median nerve, there were no difference between group A, group B and group C ( P ＞ 0.05).Conclusion After the medial anterior thoracic nerve and medial antebrachial cutaneous nerve, repaired by the contralateral C7, were severed at the beginning and at the point which was 1 cm away from the beginning of above branches, the changes of the quantity and quality of the nerve fibers from median and ulnar nerves were not significant.     

Assessment of germ cell apoptosis in cryptorchid rats

Aim: To investigate the relationship between germ cell degeneration and apoptosis in cryptorchid rats. Methods: Thirteen 21-day-old Wistar rats were made unilaterally cryptorchid by closing the left inguinal canal. At day 30 (Group 1, n=6) and day 60 (Group 2, n=7) after operation, the testes were removed for histopathological examination. The controls (n=8) were sham operated and were sacrificed at day 60. Germ cell apoptosis was assessed by means of the TUNEL method. Results: Spermatogenesis was arrested and the testicular and seminiferous tubular diameters were significantly reduced In the unilateral undescended testes (UUTs) compared with the contralateral descended testes (CDTs) and the control rats. However, atrophic changes, pathological calcification, necrosis of seminiferous tubule, and absence or sloughing of germ cells were not found in all the animals. The spermatocytes were the main type of germ cells undergoing apoptosis in all the groups. In the UUTs, there was a significant and time-depe     

Transforming growth factor β receptor Ⅱ expression inexperimental cryptorchidism and apoptosisin spermatogenic cells in rats

Objective: To study the transforming growth factor β receptor Ⅱ (TGFβ-R Ⅱ) expression in experimental cryptorchidism and apoptosis in spermatogenic cells in rats. Methods: The apoptosis of spermatogenic cells was detected by means of the terminal deoxynucleotldyl transferase mediated dUTP nick end lableling method (TUNEL) and the TGFβ-R Ⅱ expression was observed with the immunohistochemistry SABC methods. Results: There was a significant increase in the TGFβ-R Ⅱ expression in unilateral undescended testes (UUTs) compared with that in contralateral descended testes (CDTs, P<0.01). However, there was a significant and time-dependent increase in the mean apoptotic index in UUTs than in CDTs. Conclusion: TGFβ-R Ⅱ may play an important role in spermatogenic cell apoptosis.     

Effect of prolonged cryptorchidism on germ cell apoptosis and testicular sperm count

Aim:To evaluate the long term effect of experimental cryptorchidism on germ cell apoptotic rate and testicular sperm content in adult rats.Methods:Bilateral cryptorchidism was created in 40 adult male Sprague-Dawley rats by surgically manipulating the testes into the abdominal cavity and closing the internal inguinal ring.The rats were sacrificed and the testes removed 6 hours and 2,4,7,21,28 and 56 days after cryptorchidism.Germ cell apoptosis was quantified by means of TUNEL assay and apoptosis was confirmed using transmission electron microscopy.Results:The rate of apoptosis peaked at 4 days of cryptorchidism and then progressively declined to a nadir at 14 days of cryptorchidism.At 56 days of cryptorchidism,the germinal epithelium was largely depleted by the apoptotic process and only a few mature sperm were seen within the testis.At this point,a few tubules were seen to be repopulating with primary spermatocytes and the level of germ cell apoptosis began to increase marginally.Testicular sperm count (TSC) began to decline rapidly at day 7 of cryptorchidism.Only a few mature sperm were found in the testes of rats following 56 days of cryptorchidism.Multinucleated giant cells (MGC) were most numerous within the seminiferous tubules at day 4.At day 7,35% of MGCs were TUNEL positive.At all subsequent time points,however,MGCs fail to stain positive for apoptosis.This resumption of increased apoptosis coincided with the appearance of a population of primary spermatocytes in some seminiferous tubules.Moreover,there was not a corresponding increase in the number of mature sperm after 56 days of cryptorchidism.Conclusion:The decline in germ cell apoptosis after 4 days of cryptorchidism can be attributed to be the result of an overall depletion of germ cells.It appears that after a prolonged cryptorchidism (56 days),there is a limited resumption of spermatogenesis presumably as a result of a decrease in the maturing germ cells undergoing programmed cell death.(Asian JAndrol2004 Mar;6:47-51)     

Erythropoietin gene transfer into rat testes by in vivo electropo-ration may reduce the risk of germ cell loss caused by cryptorchidism

Aim： To investigate the effects of rat Erythropoietin （Epo） on spermatogenesis by transferring rat Epo gene into cryptorchid testes by means of in vivo electroporation. Methods： Sprague-Dawley rats with surgically-induced unilateral cryptorchidism were divided into three groups： the first group was given intratesticular injections of pCAGGS-Epo （pCAGGS-Epo group）, the second group was given intratesticular injections of pCAGGS （pCAGGS group）, and the third group were given intratesticular injections of phosphate-buffered saline （PBS group）. At the same time, square electric pulses of 30 V were applied six times with a time constant of 100 ms. One or two weeks after injection, each testis was weighed and the ratio of the total number of germ cells to that of Sertoli cells （G/S ratio） was calculated to evaluate the impairment of spermatogenesis. Ten testes taken from each of the three groups were examined at each time point. Results： The testicular weight after the injection of pCAGGS-Epo or pCAGGS control plasmid was （0.85 ± 0.08） g and （0.83 ± 0.03） g, respectively, at week 1 （P = 0.788） and （0.62 ± 0.06） g and （0.52 ± 0.02） g, respectively, at week 2 （P = 0.047）. At week 1, spermatids and sperm were more abundant in testes with pCAGGS-Epo than those in the control testes. At week 2, spermatids and sperm were hardly detected in either group. The G/S ratio was 23.27 ± 6.80 vs. 18.63 ± 5.30 at week 1 （P = 0.0078） and 7.16 ± 3.06 vs. 6.05 ± 1.58 at week 2 （P = 0.1471）, respectively. Conclusion： The transfer of Epo to rat testes by in vivo electroporation may reduce the risk of the germ cell loss caused by cryptorchidism.     

Cytokines in the BALB／c mouse testis in various conditions

Aim: To investigate whether testosterone, estrogens, vasectomy, experimental cryptorchidism, varicocele or aging would induce changes in the cytokine environment of the mouse testis, Methods: In adult male BALB/c mice, testosterone implants, estradiol benzoate, vasectomy, unilateral cryptorchidism, unilateral varicocele were administered/performed. The mice were followed up for different periods of time and were then sacrificed with testes incised for examination. The control mice received the vehicle or sham-operation. Results: IL-10 was present in Leydig cells of nearly every testis and IL-10 macrophages in 39% of testes. IL-6 was found in the testes of intact adult mice, mice treated with testosterone for 70 days, cryptorchid testes and sham-operated testes. Conclusion: Results suggest that IL-10 might be involved in the generation of the immunologically privileged microenvironment in the testis.     

生殖股神经在单侧隐睾鼠对侧睾丸损害中的作用机制

目的：探讨生殖股神经在单侧隐睾鼠模型对侧睾丸损害中的作用机制。方法：建立单侧隐睾鼠模型（21d龄），切断该侧生殖股神经，120d后观察对侧睾丸的生精细胞凋亡及细胞乳酸含量变化。结果：切断生殖股神经后对侧睾丸生精细胞凋亡减少，乳酸含量降低，乳酸含量与细胞凋亡呈正相关。结论：单侧隐睾症对侧睾丸损伤可能与其神经传导反射性血流减少引起生精细胞凋亡有关。     

单侧隐睾大鼠生殖股神经在对侧睾丸损害中的作用机制研究

目的 :探讨单侧隐睾大鼠模型生殖股神经在对侧睾丸损害中的作用机制。　方法 :建立单侧隐睾大鼠模型 (2 1d龄 ) ,切断该侧生殖股神经 ,12 0d后观察对侧睾丸的生精细胞凋亡变化及组织乳酸含量变化。　结果 :切断生殖股神经后 ,对侧睾丸生精细胞凋亡为 (5 .76± 0 .76 ) % ,与对照组 (17.2 8± 1.36 ) %相比明显减少 (P <0 .0 5 ) ;乳酸含量也由 (2 .19± 0 .2 4 )mmol/L下降为 (1.70± 0 .31)mmol/L(P <0 .0 5 ) ,且乳酸含量与细胞凋亡呈正相关(P <0 .0 5 )。　结论 :单侧隐睾症对侧睾丸损伤可能与其神经传导反射性血流减少引起生精细胞凋亡有关     

单侧隐睾大鼠对侧睾丸的损害与Bcl-2和Bax基因表达

目的:探讨单侧隐睾大鼠对侧睾丸生精细胞凋亡与Bcl-2/Bax基因表达的关系。方法:20只健康SD雄性大鼠(22日龄)随机分成隐睾组和对照组,每组10只。通过手术建立单侧隐睾动物模型。术后90 d取对侧睾丸,采用原位缺口末端标记(TUNEL)法检测生精细胞凋亡,免疫组化SP法检测Bcl-2/Bax基因表达。结果:与对照组相比,隐睾组对侧睾丸生精细胞凋亡显著增多(P<0.01),重量显著减轻(P<0.01),Bax表达显著升高(P<0.01),Bcl-2表达显著降低(P<0.01)。凋亡细胞主要是初级精母细胞和圆形精子细胞。结论:单侧隐睾大鼠对侧睾丸的生精细胞凋亡增多与Bcl-2基因表达降低、Bax基因表达升高密切相关。细胞内Bc l-2/Bax比值是影响生精细胞凋亡的重要因素之一。     

Pubertal genitofemoral nerve division induces testicular ascent in adult rats

BACKGROUND/PURPOSE: Ascending testes, which normally are located at the bottom of the scrotum in early infancy and later ascend back out of the scrotum, have been reported by several investigators. However, little is known about the effect of the division of the genitofemoral nerve (GFN) on testicular ascent as boys grow. The purpose of this study is to investigate whether the division of the proximal genitofemoral nerve in prepubertal rats induces testicular ascent in adulthood. METHODS: Thirty-day-old Wistar King A Rats (n = 27) underwent a unilateral proximal GFN transection on either the right or left side. At 150 days of age, the rats were killed, and their testicular position was examined. The length of the processus vaginalis was measured, and the testes were removed and weighed. Sham-operated rats were used as controls (n = 10). Student's t and the chi2 test were used for the statistical analysis. RESULTS: At 150 days of age, 21 of the 27 operated rats (77.8%) showed unilateral testicular ascent on the operated side. All testes were located at the bottom of the scrotum in sham-operated control rats (20 testes). Both the length of the processus vaginalis and the testicular weight were decreased significantly more on the operated side than in the sham-operated rats. CONCLUSIONS: These findings suggest that the proximal division of the genitofemoral nerve in prepubertal rats may induce a relative ascent of the testis by preventing the growth of the processus vaginalis in adulthood. In patients with such ascending testes, an abnormal development or accidental trauma of the genitofemoral nerve may be involved in testicular ascent.     

Does proximal genitofemoral nerve division induce testicular maldescent or ascent in the rat?

OBJECTIVE: To investigate whether the division of the proximal genitofemoral nerve (GFN) in neonatal rats induces testicular undescent or ascent in adulthood. MATERIALS AND METHODS: Neonatal Wistar King A rats underwent a unilateral proximal GFN transection on either the right or left side. At the age of 30 days, testicular descent was examined in all rats and the position of the testis recorded. The animals were allowed to develop further and the position of the testis re-examined at the age of 90-180 days, when the testes were removed and weighed. Sham-operated rats were used as controls. RESULTS: At the age of 30 days, four of the 46 (9%) operated rats showed a unilateral undescended testis on the operated side. At the age of 90-180 days, 43 rats were re-examined (three rats died before re-examination); 34 (79%) of these rats showed undescended testes on the operated side. The occurrence of cryptorchidism was significantly higher in the 90-180-day-old mature rats than in 30-day-old prepubertal rats (P<0.01). The mean (sd) weight of the undescended testes, at 2.36 (0.21) mg/g body weight, was significantly less than that of the contralateral scrotal testes, at 3.83 (0.23) mg/g; P<0.01) and of the control testes at the age of 90-180 days. In the sham-operated rats, all testes were located at the bottom of the scrotum at 30 days of age and no rats showed any testicular ascent thereafter. CONCLUSIONS: The proximal division of the GFN in neonatal rats not only causes inguinoscrotal testicular maldescent but may also induce testicular ascent in adulthood. Testicular ascent may thus be caused by some intrauterine disorders of the GFN in patients with ascending testis.     

大鼠单侧隐睾对侧睾丸的损害与抗氧化酶mRNA的表达

目的　从基因表达水平探讨单侧隐睾对侧睾丸损害机制。方法　 3 0只SD雄性大鼠分为对照组与隐睾组 ,每组各 15只。采用逆转录 聚合酶链反应 (RT PCR)方法检测对侧睾丸中谷胱甘肽过氧化物酶 (GSH PX)、铜 /锌超氧化物岐化酶 (Cu/Zn SOD)、过氧化氢酶 (CAT)mRNA的表达 ;化学比色法测定丙二醛 (MDA)的含量 ;生物素 dUTP/酶标亲和素法检测睾丸生殖细胞的凋亡。结果　术后 2周 ,与对照组比较 ,隐睾组GSH PX和SODmRNA表达明显下降 ,MDA和生殖细胞凋亡明显升高 (P <0 .0 1) ,而CATmRNA表达无明显改变 (P >0 .0 5 )。结论　单侧隐睾对侧睾丸存在GSH PX和SODmRNA表达降低 ,氧自由基升高和生殖细胞过度凋亡。     

Activation of endothelial nitric oxide synthase in contralateral testis during unilateral testicular torsion in rats

There are controversies about the injury of the contralateral testis during unilateral testicular torsion (UTT). An autonomic reflex arc between bilateral testes has been proposed. The authors focused on the involvement of nitric oxide (NO) in the contralateral testis during UTT. Eight-week-old male Wistar rats underwent unilateral torsion (1 h)-detorsion (up to 24 h). NO synthase (NOS) activity was detected as NADPH-diaphorase activity after fixation by paraformaldehyde. N-nitro-L-Arginine methyl ester (L-NAME, 20 mg/kg) was injected intravenously to the other group of rats. To evaluate the testicular injury, proteolysis of alpha-fodrin production was detected by Western blotting. Apoptosis of the germ cells was evaluated by TUNEL. Long-term effect on spermatogenesis was evaluated by flow cytometry at 60 days after UTT. Transient activation of NOS was detected following the proteolysis of alpha-fodrin in the contralateral testis. L-NAME inhibited these alterations. NADPH-diaphorase activity and eNOS immunoreactivity were co-localized in the endothelial cells. These reactions were not observed in other organs. There was neither enhanced apoptosis nor deteriorated spermatogenesis in the contralateral testis during and 60 days after UTT. In the contralateral testis, eNOS-derived NO regulates the vasomotor function against unilateral testicular torsion, whereas it acts slightly cytotoxic. These results suggest the possible involvement of a testis-specific neurovasomotor reflex between the bilateral testes.     

Overexpression of endothelial nitric oxide synthase in transgenic mice accelerates testicular germ cell apoptosis induced by experimental cryptorchidism

Surgical induction of cryptorchidism in experimental animals causes testicular germ cell apoptosis and infertility. The mechanisms of germ cell apoptosis have been associated with oxidative stress or testicular exposure to elevated temperature. Nitric oxide (NO) has been associated with apoptosis in a number of cell types. The objective of this study was to investigate whether overexpression of endothelial NO synthase (eNOS) could accelerate apoptosis of germ cells in the testes of transgenic mice. There are 3 NOS isoforms, and we restricted the analysis to eNOS at this time. For the colocalization of eNOS, staining in degenerating germ cells that were apoptotic cells suggested that eNOS may be related to germ cell apoptosis. eNOS overexpression in the testes of eNOS transgenic (eNOS-Tg) mice was examined using Western blot analysis. Unilateral cryptorchidism was surgically induced in both eNOS-Tg and wild-type (WT) adult mice. The testes were evaluated 1, 3, 5, 7, and 14 days after the operation by weighing the testes and examining histopathologic features and cell apoptosis using in situ microscopic analysis of DNA fragmentation. Immunoblotting for eNOS protein demonstrated increases in eNOS protein expression in testes, as well as the lung and aorta. In eNOS-Tg mice, weight reduction of cryptorchid testis was significantly increased on days 3, 5, and 7 (P = .02, .02, and .04, respectively). The numbers of spermatocytes and spermatids of eNOS-Tg cryptorchid testis significantly decreased compared with those of WT cryptorchid testis from day 3 (spermatocytes: P = .04; spermatids: P = .02). Moreover, terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling demonstrated that eNOS-Tg mice significantly accelerate germ cell apoptotic changes induced by experimental cryptorchidism compared with WT mice from day 3 (P = .03). We have provided evidence that eNOS plays a functional role in mouse spermatogenesis in cryptorchidism-induced apoptosis.     

大鼠一侧睾丸扭转对侧睾丸改变的实验研究

目的 :研究一侧睾丸扭转 (UTT)后对侧睾丸组织学及生精细胞凋亡的改变 ,以明确UTT后对侧睾丸是否存在损伤。　方法 :SD雄性大鼠 6 0只 ,随机分为实验组 (n =4 8)及对照组 (n =12 )。实验组采用Turner方法建立左侧睾丸扭转模型 ,于扭转后 6h处死 4只 ,其余 4 4只再分为扭转睾丸复位及切除组 ,分别于术后 1d、1周、4周处死7～ 8只 ,取睾丸组织进行组织学及生精细胞凋亡的检测。　结果 :UTT复位后对侧睾丸组织学发生明显改变 ,生精细胞凋亡指数明显高于对照组 (P <0 .0 5 )。扭转睾丸切除后对侧睾丸变化不明显。　结论 :UTT可引起对侧睾丸损伤 ,其机制可能与再灌注有关 ,扭转睾丸切除可防止或减轻对侧睾丸的损伤