Protection of spermatogenesis against cytotoxic effects of two chemotherapeutic drugs by temporary testicular blood flow interruption in the ram

Summary. Temporary interruption of the testicular blood flow for 1 h after injection of cytostatic drugs has a protective effect on spermatogenesis. This was shown in experiments in which spermatogenesis was evaluated at four weeks after a single intravenous injection of Adriblastina^R (ADR; doxorubicine hydrochloride) or Mitomycin-C-kyowa^R (MIT). Interruption of the blood flow was performed by inflation of an occluder implanted around the testicular artery.
The animals were killed and histological sections prepared 4 weeks after treatment. In all drug-treated animals spermatids were near absence and spermatocytes were decreased in number. Therefore, even after occlusion of the blood flow, the drug doses were high enough to kill not only large numbers of differentiating spermatogonia but also stem cells.
The response of the stem cells to the treatments was evaluated by counting the numbers of A spermatogonia per 100 Sertoli cells in the different groups. Normal numbers of these cells were found after both MIT and ADR, indicating that the stem cell population had responded to the initial cell loss by extra proliferation. However, significantly higher numbers of A spermatogonia were found in the drug-treated animals in which the testicular blood flow was interrupted for 1 h. This indicates that occlusion of the blood flow to the testis for 1 h results in a faster recovery of spermatogenesis than after drug treatment alone.     

Effects of local heating of the testis on testicular blood flow and testosterone secretion in the rat

Exposure of one or both testes of rats to heating at 43 degrees C for 30 min resulted in a significant reduction in blood flow per testis, as measured using microspheres. The effects on the testes of unilateral and bilateral heating were similar, although the changes in FSH levels in peripheral blood were in general less marked after unilateral heating. Testicular blood flow fell, along with testicular weight, beginning at 2-4 days and reaching minimum values 14-21 days after heating. Both blood flow per testis and testicular weight were beginning to recover 35 days post-heating and blood flow per testis was normal by 56 days following heat treatment, although testicular weight was still slightly reduced at that time. Heating one or both testes to 42 degrees C produced similar but smaller responses 21 days later, whereas temperatures of 41 degrees C or lower were without effect on the parameters measured, except for some rises in serum LH and FSH. With slight reductions in blood flow, there were corresponding increases in testicular venous testosterone concentration so that testosterone secretion was unaffected. Further reductions in blood flow at 14 and 21 days after heating to 43 degrees C were not fully compensated by an increase in the concentration of testosterone in testicular venous blood, with the result that testosterone secretion fell.     

Arginine-vasopressin induced changes in testicular blood flow

As arginine-vasopressin (AVP) has been suggested as a possible paracrine regulator in the testis, the present study was designed to evaluate the effects of AVP on testicular blood flow and vascular permeability. Intratesticular injection of AVP caused a dose-dependent decrease in total testicular blood flow, with 1 ng AVP being the lowest dose that induced a decrease. The AVP-induced decrease in blood flow was blocked completely by treatment with an AVP antagonist, but the latter had no effect on its own. High doses of AVP (100 and 1000 ng) decreased testicular IF volume and slightly increased the IF testosterone concentration. At 10 min and 1 h after intratesticular injection of 100 ng AVP testicular blood flow was decreased, followed by an increase at 6 h; testosterone concentrations were increased at both 10 min and 6 h after this treatment. The microcirculatory blood flow pattern (vasomotion) was not influenced by AVP. The present study shows that AVP in low doses influences testicular blood flow rather selectively because AVP was without any major effect on vasomotion, interstitial fluid volume and testosterone production. AVP might therefore be of importance in the regulation of testicular blood flow.     

Exogenous oxytocin administration improves the testicular blood flow in rams

The current research aimed to examine the effects of single-dose oxytocin administration on testicular blood flow measurements peak systolic velocity (PSV), end-diastolic velocity (EDV), resistive index (RI) and pulsatility index (PI) and plasma steroid (testosterone and oestradiol-17β) concentrations in rams. In the treated group, eight mature Ossimi rams during the breeding season were injected intravenously (iv) with 20 IU oxytocin, while the other eight male rams were administered normal saline (2 ml) iv as a control group. Venous blood samples and testicular blood flow in the left and right testes were examined immediately before (0) and 5, 30, 60 and 120 min after injections. The control group did not show significant changes in the variables examined (p > .05), except for the EDV (p < .05). In the treated group, the administration of oxytocin led to a significant decrease in RI and PI values starting 5 min after oxytocin administration until 60 min after its application (p < .05). The plasma concentrations of testosterone and oestradiol-17β raised from 5 to 30 min after oxytocin injection (p > .05) and then declined significantly until 120 min (p < .05). In conclusion, oxytocin is a potent testicular vasodilator affecting the testicular vascular tone and steroid concentrations in rams.     

Stimulatory effect of an LHRH-agonist on testicular blood flow in hypophysectomized rats

The effect of an LHRH-agonist (A) on testicular blood flow and plasma testosterone levels was studied in adult hypophysectomized rats. Subcutaneous injection of 1 microgram LHRH-agonist 2 or 4 h prior to experiment induced an increase in testicular blood flow. (Controls: 21.1 +/- 2.8 ml/100 g X min; LHRH-A + 2 h: 28.9 +/- 3.3 ml/100 g X min; LHRH-A + 4 h: 32.4 +/- 4.1 ml/100 g X min P less than 0.05; all mean +/- SEM). There was also a significant increase in plasma testosterone levels at 2 h (P less than 0.01) and 4 h (P less than 0.01) after treatment with the LHRH-agonist, with a peak at 2 h.     

Unilateral spermatic cord torsion without ipsilateral spermatogenetic material: effects on testicular blood flow and fertility potential

This experiment was planned to answer the question of how the elimination of ipsilateral spermatogenetic material, which is necessary for contralateral testicular damage caused by an autoimmune response, affects contralateral testicular blood flow and fertility potential in unilateral spermatic cord torsion (USCT). Thirty-four male and 68 female adult albino rats were divided into three groups. Group 1 rats underwent a control operation, group 2 rats underwent subepididymal orchiectomy to eliminate spermatogenetic material, and group 3 rats underwent USCT after subepididymal orchiectomy. Testicular blood flows of the rats were measured by ¹³³Xe clearance technique. Additionally, to determine fertility potential, each male rat was housed with two female rats. Numbers of impregnated and delivered rats were recorded. Both mean testicular blood flow and fecundity of group 3 were significantly lower than those of groups 1 and 2. When compared with groups 1 and 2, fertility and mean number of the impregnated rats of group 3 were lower but the differences were not significant. These findings suggest that absence of spermatogenetic material in USCT reduces contralateral blood flow and fertility potential. Therefore, contralateral testicular damage originating from blood flow alterations rather than autoimmune mechanism should be considered to explain fertility problems encountered following USCT.     

The early effects of sympathectomy on bone blood flow

Summary Several laboratories, including our own, have investigated the physiology of bone blood flow by perfusion of the tibia via the nutrient artery. Any damage caused to the nerve supply by cannulation of the artery might affect the results. The possible effect of such damage was investigated by measuring blood flow to the dog tibia using a microsphere technique, before and after sympathectomy. No effect was found and the tibial nutrient artery is therefore a suitable vessel for perfusion when estimating nutrient exchange in bone.     

The effects of increased intraabdominal pressure on testicular blood flow, oxidative stress markers, and morphology

Background

This study was carried out to evaluate the effects of increased intraabdominal pressure (IAP) on testicular blood flow (TBF), oxidative stress markers, and morphology.

Methods

Twenty-four Sprague-Dawley rats weighing 300 to 350 g were allocated randomly into 3 groups consisting of 8 animals each: A, gasless (control); B, 10 mm Hg IAP with CO₂ pneumoperitoneum for 60 minutes; and C, 20 mm Hg IAP with CO₂ pneumoperitoneum for 60 minutes. Testicular blood flow was studied using the Doppler technique. In the 10 and 20 mm Hg IAP groups, time points of TBF measurements were defined as follows: TBF_baseline, 10 minutes before insufflation; TBF_10min, 10 minutes after pneumoperitoneum; TBF_50min, 50 minutes after pneumoperitoneum; and TBF_reperfusion, 10 minutes after pneumoperitoneum deflation. To evaluate the changes in oxidative stress, we assayed the malondialdehyde (MDA) levels of testicular tissues. A 4-level grading scale was used to quantify histologic injury.

Results

For both testes of each rat, TBF_10min, TBF_50min, and TBF_reperfusion values of each group were separately evaluated according to their TBF_baseline value percentages. The results revealed no significant differences for each time point of TBF measurements between the right and left testes in any group. Pneumoperitoneum caused a significant decrease in TBF at the 10th and 50th minutes of pneumoperitoneum, both in the 10 and 20 mm Hg IAP groups, compared with their baseline values. TBF_reperfusion values in both groups were also lower than their baseline values. We determined that mean TBF_10min and TBF_50min values decreased significantly in the 20 mm Hg IAP group compared with the 10 mm Hg IAP group, despite there being no significant difference in their mean TBF_reperfusion values.Mean MDA levels were significantly increased in both the 10 and 20 mm Hg IAP groups compared with those of the control group for the right and left testes. However, there was no significant difference between the mean MDA levels in these first 2 groups. The histologic injury score was significantly increased in both the 10 and 20 mm Hg IAP groups compared with the control group; however, there was no difference in the scores between these first 2 groups.

Conclusions

We demonstrated in an animal model that abdominal deflation after IAP of 10 and 20 mm Hg for 60 minutes causes testicular hypoperfusion, free radical production, and subsequent testicular damage.     

The effect of melatonin implants on blood testosterone and acrosin activity in spermatozoa of the ram

In a series of consecutive blood sampling in 15 days intervals over 15 weeks after implantation of melatonin in rams an increased mean value, basal level and number of peaks of testosterone was observed in samples of the third fortnight (45th day). This increase was greater in the autumn (breeding season) than in spring (non-breeding season). Total acrosin activity in spermatozoa was increased between days 35-56 (autumn) and days 49-70 (spring) after implantation and the relative increase was higher in autumn than in spring. The increase of acrosin activity was independent of the changes of testosterone. An increase of acrosin activity by melatonin, in cases of low activity, might improve fertilization rates in sheep not only during the breeding season, but also during the non-breeding season (after oestrus induction).     

Does follicle-stimulating hormone or pregnant mare serum gonadotrophin influence testicular blood flow in rats?

Treatment of adult rats with 25 iu follicle-stimulating hormone (FSH) did not influence testicular blood flow, serum testosterone, vasomotion or intravascular leucocyte concentration at 6, 12 and 24 h after treatment. Treatment of adult rats with 50 iu pregnant mare serum gonadotrophin (PMSG) resulted in a two-fold increase in testicular blood flow at 24 h, and an increase in interstitial fluid volume at 36 h after treatment. This PMSG-induced increase in blood flow did not occur in Leydig cell-depleted animals, suggesting that the effect on blood flow is mediated via the Leydig cells. PMSG injection stimulated testosterone secretion but it did not influence vasomotion, and it only marginally increased the secretion of leucotactic factors in the testis. The present study suggests that FSH has no apparent effects on testicular blood flow and that the effects of PMSG (a hormone with both FSH and LH-like activity) is mediated via stimulation of the Leydig cells.     

腹腔镜完全腹膜外疝修补术对睾丸血流及血清睾酮的影响

【摘要】〓目的〓初步探讨腹腔镜完全腹膜外无张力腹股沟疝修补术(TEP)对睾丸血流及血清睾酮的影响。方法〓对我科住院行手术治疗的男性单侧腹股沟疝患者进行前瞻性研究,应用高频彩超对行腹腔镜下腹股沟疝修补的男性患者在术前48 h内,术后48 h内、2周内、1个月内用彩色超声分别测定患者患侧和对侧精索动脉血流量,包括收缩期峰值血流速度(PSV)和舒张末期血流速度(EDV)和睾丸体积,以及检测血清睾酮水平。结果〓共66例最终被选择入组。术前48 h内,患侧和对侧的PSV、EDV和睾丸体积差异均无明显异常(P>0.05);术后48 h及术后两周均检测到患侧的PSV、EDV和睾丸体积明显小于对侧,差异具有统计学意义(P<0.05)。而术后1个月内,患侧和对侧的PSV、EDV和睾丸体积差异均无统计学意义(P>0.05)。术前48 h内,术后48 h内、2周内及1个月的患者血浆睾酮水平的差异无统计学意义(P>0.05)。结论〓TEP术治疗腹股沟疝时,术后早期(2周内)可降低患侧的睾丸血流和睾丸体积,但术后1个月可恢复正常;TEP术对腹股沟疝患者的血清睾酮无明显影响。不需进行干预。     

A review of the effects of external pressure on skin blood flow

The human foot is a complex mechanical structure consisting of bones, ligaments and joints. They act together to provide a robust system capable of absorbing and dissipating the intermitted pressure that is subjected to its plantar surface during walking to prevent soft tissue breakdown. Current studies suggest that plantar foot pressure may lead to soft tissue breakdown (e.g. neuropathic ulceration) and hence research has so far concentrated on investigating the mechanical effects of plantar foot pressure on the foot’s integrity. This has been possible through the widely available pressure and force platforms as well as in-shoe pressure systems. However, to understand how plantar foot pressure causes soft tissue breakdown it is vital to investigate both the physiological–mechanical interactions between the skin and plantar foot pressure. This review suggests that with the current advances in technology, the physiological response of skin blood flow to mechanical plantar foot pressure should be investigated and correlated further, both during static and dynamic loading, by developing a new system capable of either measuring both variables simultaneously or by synchronising two systems in real time.     

The effects of local and sustained release of fibroblast growth factor on testicular blood flow and morphology in spermatic artery--and vein-ligated rats

Background/purpose

This study was carried out to evaluate the effects of local and sustained release of fibroblast growth factor (FGF) on testicular blood flow and morphology in spermatic artery— and vein-ligated rats.

Methods

Forty male Wistar albino rats weighting 300 ± 20 g were allocated randomly into 5 groups consisting of 8 in each as follows: G-S (sham); G-C (control); and G-T_0.85, G-T_1.70, G-T_2.55. After the ligation of the left spermatic artery and vein, 1 cm² of unloaded and 0.85 μg, 1.70 μg, and 2.55 μg of FGF-loaded gelatin films were sutured on the left epididymis in G-C, G-T_0.85, G-T_1.70, and G-T_2.55, respectively. After 30 days, bilateral capsular (CBF) and intratesticular (IBF) blood flows were evaluated by colored Doppler ultrasonography (CDUS) and testicular blood flow (TBF) by ¹³³Xe clearance technique. Tunica albuginea and intertubular tissues were studied for the increase of peritesticular and intratesticular vessels. Mean intertubular vascular structure counts, seminiferous tubular diameters, testicular biopsy scores, and Leyding cell scores of each group were recorded and compared.

Results

CBF was present in all animals of G-S, G-T_0.85, G-T_1.70, and G-T_2.55 groups in CDUS, and it was detected in 62% of the G-C rats (P < .05). However, IBF was present in only 25% of the G-C rats, and this percentage was increased from 50% up to 87.5% for treatment groups, and 100% for G-S rats, respectively. ¹³³Xe clearance showed that TBF was significantly decreased in G-C compared with G-S (P < .05). In G-T_2.55, TBF was significantly increased, but still could not reach the level of G-S. Although mean testicular weights were significantly decreased for controls (G-C), G-T_0.85, and G-T_1.70, almost no difference was observed between G-T_2.55 and G-S. Although a slight increase in the vascular structures of tunica albuginea was present in G-C rats, a significant increase was observed in treatment groups. The mean number of intertubular vascular structures was significantly increased in treatment groups when compared with G-S and G-C (P < .05). Mean seminiferous tubular diameters and Leydig cell scores were decreased in G-C but significantly increased in treatment groups (P < .05). Mean testicular biopsy scores were increased in treatment groups compared with G-C but could not reach to sham levels.

Conclusions

Ligation of the spermatic artery and vein has detrimental effects on the ipsilateral testicular blood flow and morphology. These effects may be reversed by local application of FGF.     

Pressure flow pattern of varicocele veins and its correlation with testicular blood flow and semen parameters

The pressure pattern in varicocele veins of infertile patients and its correlation with semen quality and testicular blood flow was determined. Consecutive patients at andro‐urology clinic of a teaching hospital undergoing microsurgical varicocelectomy were included. Their semen quality and testicular blood flow were determined. Peak systolic velocity (PSV) and resistive index (RI) of subcapsular and intraparenchymal branches of testicular artery were noted by colour Doppler ultrasonography. During surgery before ligation of varicocele veins, intravenous pressures of internal spermatic (ISV) and external spermatic (ESV) veins were determined at baseline and after Valsalva manoeuvre. Thirty patients, 20–45 years old, were evaluated. Baseline pressure for maximum dilated ISV (A), less dilated ISV (B) and ESV was 15.93 ± 6.34, 12.38 ± 4.60 and 12.92 ± 5.65 mm. Hg, respectively, which increased after Valsalva by 104.4%, 116.2% and 38.22% respectively. Correlation (r = ?.71; p < .05) was appreciated between percentage increase in pressure of ISV B with PSV of intraparenchymal testicular arteries and progressive motility (r = ?.759; p < .05), nonprogressive motility (r = ?.738; p < .05) and morphology (r = ?.653; p = .07) of spermatozoa. In conclusion, ISV develops higher pressure on Valsalva as compared to ESV and has correlation with semen quality and testicular blood flow.     

Experimental studies of the effects of local hyperthermia on blood flow,oxygen pressure and pH in tumors

The heat-induced environmental changes in tumor tissues are considered to influence the antitumor effect of hyperthermia or hyperthermochemotherapy, which is believed to complement the direct lethal effect of heat on tumor cells. The effects of local hyperthermia on the blood flow, oxygen pressure and pH in tissues were investigated using AH-100B tumor bearing rats, by immersing the tumor in a water bath at 41°, 43° and 45°C. These parameters were measured in the marginal and deeper sites of the tumor mass, and in the normal muscle adjacent to the tumor. During immersion at 41°C, blood flow in the tissue was increased at each site, and during immersion at 43°C, tissue blood flow increased initially at each site, but decreased with time to rates below that of the unheated tissue. During immersion at 45°C, the blood flow decreased markedly in each tissue. The changes in oxygen pressure and pH in each tissue were similar to those observed in the blood flow during localized heating at 41°, 43° and 45°C. In local thermochemotherapy, the initial stage of hyperthermic treatment seems to be the most suitable time for administering carcinostatics, since it is the time when tumor blood flow has not yet decreased.     

Collection of peripheral blood stem cells with granulocyte-colony-stimulating factor alone in testicular cancer patients.

BACKGROUND: High-dose chemotherapy with the transplantation of peripheral blood stem cells (PBSC) has been performed for the treatment of advanced testicular cancer patients. Recently, it has been reported that, in healthy donors, a large quantity of stem cells can be transferred to peripheral blood using granulocyte-colony-stimulating factor (G-CSF) alone. Therefore, it was decided to try to harvest PBSC from three patients having testicular cancers with G-CSF alone. METHODS: The three patients with testicular cancer were 26, 56 and 62-years-old. They had undergone five, two and three cycles of chemotherapy, respectively, but no radiation therapy. Granulocyte colony-stimulating factor was subcutaneously injected (250 microg) into each patient twice per day for 6 days. Peripheral blood stem cells were harvested for 3 days (days 4-6) and mononuclear cells (MNC), CD34-positive cells and colony-forming units of granulocyte-macrophage (CFU-GM) in PBSC collected by apheresis were measured. RESULTS: Apheresis showed that the total MNC count was 20.2 x 10(8)/kg (range, 10.6-25.9 x 10(8)/kg), the CD34-positive cell count was 0.98 x 10(6)/kg (range, 0.75-1.4 x 10(6)/kg) and the total CFU-GM count was 1.36 x 10(5)/kg (range, 0.25-3.0 x 10(5)/kg). CONCLUSION: After mobilization of peripheral blood stem cells with G-CSF alone, sufficient amounts of MNC were obtained from testicular cancer patients who had undergone chemotherapy several times. However, sufficient amounts of CD34-positive cells and CFU-GM could not be obtained. These results suggested that the G-CSF dose was not adequate for harvesting sufficient amounts of CD34-positive cells and CFU-GM.     

选择性半肝血流阻断对结直肠癌肝转移切除术后疗效的影响

目的 研究选择性半肝血流阻断方法对结直肠癌肝转移切除术后疗效的影响.方法 回顾性分析1998-2006年海军总医院对71例结直肠癌肝转移病人实施手术切除的情况,比较半肝血流阻断和Pringle手法两种不同的入肝血流阻断方法对于肝转移癌的治疗效果.结果 全组病人术后1,3,5年生存率为82.7%,46.2%,27.1%.采用半肝血流阻断的方法肝转移癌切除术病人的1,3,5年生存率为89.7%,54.2%,34.1%,采用Pringle手法肝门血流阻断的方法行肝转移癌切除术病人的1,3,5年分别为73.3%,41.5%,23.9%.两组1,3,5年生存率比较有显著性差异(P<0.05).结论 与Pringle手法相比较,利用半肝血流阻断的方法行肝转移癌切除术可以有效延长病人的生存期,半肝阻断方法行转移癌切除术,是减少循环肿瘤细胞的种植与减缓生长速度的有效方法.     

选择性半肝血流阻断对结直肠癌肝转移切除术后疗效的影响

目的 研究选择性半肝血流阻断方法对结直肠癌肝转移切除术后疗效的影响.方法 回顾性分析1998-2006年海军总医院对71例结直肠癌肝转移病人实施手术切除的情况,比较半肝血流阻断和Pringle手法两种不同的入肝血流阻断方法对于肝转移癌的治疗效果.结果 全组病人术后1,3,5年生存率为82.7%,46.2%,27.1%.采用半肝血流阻断的方法肝转移癌切除术病人的1,3,5年生存率为89.7%,54.2%,34.1%,采用Pringle手法肝门血流阻断的方法行肝转移癌切除术病人的1,3,5年分别为73.3%,41.5%,23.9%.两组1,3,5年生存率比较有显著性差异(P<0.05).结论 与Pringle手法相比较,利用半肝血流阻断的方法行肝转移癌切除术可以有效延长病人的生存期,半肝阻断方法行转移癌切除术,是减少循环肿瘤细胞的种植与减缓生长速度的有效方法.     

选择性半肝血流阻断对结直肠癌肝转移切除术后疗效的影响

目的 研究选择性半肝血流阻断方法对结直肠癌肝转移切除术后疗效的影响.方法 回顾性分析1998-2006年海军总医院对71例结直肠癌肝转移病人实施手术切除的情况,比较半肝血流阻断和Pringle手法两种不同的入肝血流阻断方法对于肝转移癌的治疗效果.结果 全组病人术后1,3,5年生存率为82.7%,46.2%,27.1%.采用半肝血流阻断的方法肝转移癌切除术病人的1,3,5年生存率为89.7%,54.2%,34.1%,采用Pringle手法肝门血流阻断的方法行肝转移癌切除术病人的1,3,5年分别为73.3%,41.5%,23.9%.两组1,3,5年生存率比较有显著性差异(P<0.05).结论 与Pringle手法相比较,利用半肝血流阻断的方法行肝转移癌切除术可以有效延长病人的生存期,半肝阻断方法行转移癌切除术,是减少循环肿瘤细胞的种植与减缓生长速度的有效方法.