睾丸扭转后生精细胞凋亡与iBOS基因表达

本文研究了睾丸扭转复位后生精细胞凋亡与iBOS基因表达的关系。采用大鼠建立左侧睾丸扭转复位模型（720,2h)。用TUNEL法和免疫组化SP法分别检测扭转复位后第五天生精细胞凋亡和iBOS基因表达。研究发现凋亡主要见于染色质降解的生精细胞（初级精母细胞和圆形精子细胞）。间质细胞和支持细胞未见凋亡发生。iBOS表达见于各级生精细胞,在染色质降解的生精细胞（即凋亡细胞）强表达。本文研究表明睾丸扭转复位后生精细胞凋亡增加与iBOS基因表达密切相关。睾丸局部NO生成异常可能是生精细胞凋亡增加的原因之一。     

睾丸扭转后生精细胞凋亡与iNOS基因表达

本文研究了睾丸扭转复位后生精细胞凋亡与iNOS基因表达的关系。采用大鼠建立左侧睾丸扭转复位模型(720,2h)。用TUNEL法和免疫组化SP法分别检测扭转复位后第五天生精细胞凋亡和iNOS基因表达。研究发现凋亡主要见于染色质降解的生精细胞(初级精母细胞和圆形精子细胞)。问质细胞和支持细胞未见凋亡发生。iNOS表达见于各级生精细胞,在染色质降解的生精细胞(即凋亡细胞)强表达。本研究表明睾丸扭转复位后生精细胞凋亡增加与iNOS基因表达密切相关。睾丸局部NO生成异常可能是生精细胞凋亡增加的原因之一。     

Protective role of natural antioxidant supplementation on testicular tissue after testicular torsion and detorsion

OBJECTIVES: To investigate the impact of garlic extract (GE), which is known for its antioxidant activity, on a testicular torsion/detorsion model in animals and to help understand how to prevent both ischemic and reperfusion injuries after testicular torsion and detorsion. MATERIAL AND METHODS: Six groups of rats (n=7 in each group) were used. The animals in the control group (Group I) did not receive any treatment. The animals in the sham group (Group II) underwent scrotal incision and testicular fixation only. The animals in Groups III-VI underwent 720 degrees of left testicular torsion for 2 h; subsequent detorsion was performed for 2h in Groups IV and VI only. Animals in Groups V and VI were treated exactly the same as those in Groups III and IV, respectively except that they were pretreated with oral GE for 5 days at a dosage of 5 ml/kg. Both testicles in all rats were removed and tissue malondialdehyde (MDA) levels and enzymatic activities of xanthine oxidase (XO) were studied, in addition to a histological evaluation after hematoxylin-eosin staining. RESULTS: Testicular MDA levels and XO activities were higher in Group III compared to Group II (p<0.05). Pretreatment with GE prevented these increases. Detorsion caused more damage and resulted in a further increase in MDA levels but MDA levels were not increased in animals pretreated with GE. Histologically, torsion caused some separation between germinative cells in the seminiferous tubules, which became much more prominent in Group IV and was attenuated by GE pretreatment. There were no significant changes in any of the above-mentioned enzymatic activities or histopathologic changes in the contralateral testicle in any of the groups. CONCLUSIONS: We believe that both testicular torsion and detorsion result in testicular tissue damage by means of lipid peroxidation, which is evident by an increase in the tissue levels of MDA. Dietary supplementation with GE seems to attenuate the generation of toxic free radicals, as evidenced indirectly by low tissue MDA levels.     

地塞米松对大鼠睾丸扭转复位后保护作用的研究

目的探讨地塞米松对大鼠睾丸扭转复位后的保护作用.方法24只成年健康SD雄性大鼠随机分为3组A组为睾丸扭转复位加生理盐水组,B组为睾丸扭转复位加地塞米松组,C组为对照组.建立单侧睾丸扭转复位模型.术后24h取手术侧睾丸,化学比色法测定睾丸组织的SOD、CAT、MPO活性和MDA含量,光镜观察病理学变化.结果与A组相比,B组的MDA含量和MPO活性降低(P＜0.01),SOD和CAT的活性增高(P＜0.01),睾丸被膜下白细胞浸润减少,无间质水肿.结论地塞米松可减轻大鼠睾丸扭转复位后的再灌注损伤,对睾丸有保护作用.     

Method for intraoperative assessment of organ perfusion and viability using a miniature oxygen sensor

A specially designed miniaturized Clark polarographic electrode was used to measure organ surface oxygen tension during surgical operations in 10 patients as a means of determining tissue perfusion and viability. When applied to organ surfaces, the sensor noninvasively provides real-time assessment of tissue PO2. Values obtained are dependent on both arterial PO2 and local blood flow and therefore may be used to quantitatively assess local oxygen delivery (perfusion). Measured values may be compared with arterial PO2 or to areas of certain normal perfusion on the same organ to better define the adequacy of local oxygen transport. Tissue temperature is independently measured and may be used to assess the metabolic activity of the tissue monitored. Organ surface oximetry using the miniature PO2 sensor is a practical and repeatable method for the intraoperative assessment of organ perfusion and viability.     

Pulse oximetry: a new non-invasive assessment of peripheral arterial occlusive disease

Peripheral skin perfusion reflects the level of vascularity and viability of a limb and may help in planning the site of amputation or bypass surgery in patients with vascular disease. This study used peripheral pulse oximetry in 20 healthy volunteers and in 20 patients with limb ischaemia. Pulse oximetry saturation levels (Psa,O2) were compared with ankle artery Doppler pressures and transcutaneous oxygen measurements (Ptc,O2). Recordings were taken at two standard sites distally and referenced to finger and forearm to calculate an index. A significant correlation was found between Ptc,O2 and Psa,O2 in patients with ischaemia (r = 0.68, P less than 0.01). A further group of 12 patients with acute limb ischaemia was similarly assessed before and after revascularization. After revascularization mean(s.d.) Ptc,O2 increased from 38(13) to 44(1) mmHg (P greater than 0.05) and mean(s.d.) Psa,O2 increased from 86(3) to 90(4) per cent (P less than 0.01). These data suggest that pulse oximetry is a more sensitive index of peripheral perfusion than Ptc,O2 or ankle artery Doppler pressure and that, because of its accuracy and simplicity, it merits further use.     

Co-administration of Salvia miltiorrhiza and verapamil inhibits detrimental effects of torsion/detorsion on testicular tissue in rats

Testicular torsion/detorsion is one of the important emergencies that requires fast surgical intervention. This study aimed to investigate the effects of Salvia miltiorrhiza hydroalcoholic extract combined with verapamil on testicular ischaemia/reperfusion damage in Wistar albino rats. All animals were distributed in 3 groups (n = 8), including the sham-operated group, torsion/detorsion (TD) group and torsion/detorsion + pretreatment with 200 mg/kg Salvia miltiorrhiza extract combined with 0.3 mg/kg verapamil (SMV) group. Oxidative stress biomarkers (MDA, GPx, CAT and TAC) both in plasma and testicular tissue, sperm parameters (motility, vitality, concentration and morphology) and histopathological parameters (MSTD, GECT, Johnson's score, Cosentino's score and testicular cell thickness) were assessed in all groups. Ischaemia/reperfusion significantly increased MDA and decreased GPx, CAT and TAC levels (p < .05). Pretreatment with SMV significantly increased GPx, CAT and TAC levels (p < .05). SMV group increased progressive sperm motility and vitality and reduced non-progressive motility of spermatozoon (p < .05). Testicular torsion significantly decreased all histopathological parameters compared to the sham group (p < .05). SMV pretreatment remarkably increased MSTD, GECT and Cosentino's score in comparison with the TD group (p < .05). A combination of Salvia miltiorrhiza with verapamil could reduce damages triggered by testicular torsion detorsion and improve sperm functionality parameters and oxidative stress defence systems.     

“生精散”对大鼠睾丸扭转复位后生精功能的影响及其机制研究

目的:研究"生精散"对大鼠睾丸扭转复位后生精功能的影响及其机制。方法:40只雄性SD大鼠随机分为假手术组(A组),对照组(B组),生精散组小(C组)、中(D组)、大剂量组(E组),每组8只。建立左侧睾丸扭转模型,B组自扭转前1 h开始给予每日灌胃生理盐水1 ml/d,C组(0.01 g/kg.d)、D组(0.02 g/kg.d)、E组(0.03 g/kg.d)分别按体重给予灌服生精散,连续35 d后处死大鼠,对大鼠进行精液常规分析,用RT-PCR检测大鼠精子CatSper1的表达。结果:a+b级精子百分率、精子存活率、精子浓度,CatSper1基因表达,与A组[(51.30±6.60)%、(69.01±7.20)%、(40.53±7.01)×106/ml、2.04±0.77]相比,B组[(15.30±6.30)%、(44.42±6.36)%、(21.00±6.14)×106/ml、1.12±0.50),均显著降低(P<0.01);与B组相比,D组[(51.63±3.20)%、(72.09±2.20)%、(55.30±5.90)×106/ml、2.11±0.20]、E组[(55.93±3.17)%、(73.01±2.11)%、(58.33±4.90)×106/ml、2.31±0.17]均显著升高(P<0.01),而C组[(18.02±0.23)%、(48.04±7.01)%、(22.87±2.10)106/ml、1.19±0.51]升高不明显(P>0.05)。结论:生精散可以促进睾丸扭转复位后精子质量的恢复,其机制可能与调节生精功能,提高精子细胞CatSper1基因的表达有关。     

Ex vivo perfusion for accurate assessment of liver graft viability in dogs

A new type of ex vivo liver perfusion model (EVPM) was developed. The system includes a physiological porto-portal connection, using a centrifugal pump and an aorto-aortal connection, through a heparin-coated tube. Ten pairs of mongrel dogs, weighing 13-20 kg, were used. Grafts of the liver procured from donors were perfused on the EVPM for 3 h. These grafts were divided into two groups: group A (n = 5), nonpreserved graft; group B (n = 5), 8-h graft preserved with lactated Ringer's solution (4 degrees C). The orthotopic liver transplantation (OLTx) series with the nonpreserved graft was reviewed, and dogs surviving for over 7 days following OLTx, group C (n = 5), were selected for liver grafts in the actual OLTx. In comparison with groups A and C, no difference was noted among enzyme levels (sGOT, sGPT, LDH) or in the recovery rate of ATP content in the graft liver tissue after revascularization. There were significant differences between group A and group B after revascularization. This simple and physiological EVPM accurately reflects graft function and hemodynamics in the actual OLTx. Graft viability, as a substitute for OLTx, in large animals can be reliably assessed. This EVPM is expected to contribute to research on events related to liver transplantation.     

Value of an intraoperative real time tissue perfusion assessment system following a nipple-sparing radical mastectomy for advanced breast cancer

INTRODUCTION

Standard surgical approach for advanced breast cancer is a modified radical mastectomy with a periareolar elliptical incision. Here a unique surgical approach is presented utilizing intraoperative real time tissue perfusion technology.

PRESENTATION OF CASE

A 65-year old African American female was diagnosed with grade 3 papillary carcinoma on biopsy of a palpable 3.7 cm left breast mass at 12 o’clock position. Pathology showed a T2N0M0 invasive ductal carcinoma, triple negative disease, with clear margins. Patient opted for naturopathic treatment. Six months later, cancer recurred at the local site; the patient persisted with natural remedies. Nine months later, the mass had progressed to 14.5 cm with smaller adjacent nodules and nodular thickening of breast with no metastatic disease. Patient received neoadjuvant chemoradiation. Ten months later, patient underwent a nipple-sparing radical mastectomy given the persistent pectoral muscle involvement of this mass.

DISCUSSION

An elliptical incision was made in the superior aspect of the breast wall at least 6 cm above the areola. After tumor removal, the skin edges were approximated. It was decided preoperatively to use the SPY Elite™ system to assess perfusion of the flap edges. Angiography with indocyanine green showed poor perfusion of the inferior flap and an additional 2 cm of skin was resected. Final pathology showed that all margins were clear of disease and patient recovered well from surgery.

CONCLUSION

This case study shows that an individualized approach resulted in an optimal outcome for the patient. The infrequent procedure of a nipple-sparing radical mastectomy was done in coordination with SPY Elite™ to achieve good wound healing.     

Testicular torsion–detorsion and potential therapeutic treatments: A possible role for ischemic postconditioning

Testicular torsion is a common urological emergency among adolescent boys and young men. Rotation of the testis and twisting of the spermatic cord rapidly leads to ischemia, resulting in a loss of germ cells. Thus, prompt diagnosis and urgent surgical intervention are required, but the subsequent release of the torsion induces reperfusion injury, which causes further damage to the ischemic testis. Testicular torsion–detorsion (ischemia–reperfusion) injury triggers the generation of reactive oxygen species, pro‐inflammatory cytokines, neutrophil recruitment, lipid peroxidation, anoxia and apoptosis, which carry a significant risk of subsequent infertility. Previously, the effects of numerous pharmacological agents and treatments have been evaluated to prevent testicular ischemia–reperfusion injury in animal models. We propose a new treatment, especially postconditioning, to prevent adverse effects of ischemia–reperfusion injury after testicular torsion–detorsion.     

The protective role of caffeic acid phenethyl ester (CAPE) on testicular tissue after testicular torsion and detorsion

Testicular artery occlusion causes an enhanced formation of reactive oxygen species, which contributes to the pathophysiology of tissue damage. Here, we have investigated the effects of caffeic acid phenethyl ester (CAPE), a new antioxidant and antiinflammatory agent, in rats subjected to testicular torsion/detorsion (T/D). Thirty-five male rats were divided into four groups: sham operation group (n=8), torsion group (n=9), T/D+saline group (n=9) and T/D+CAPE group (n=9). Rats, except the sham operation group, were subjected to left unilateral torsion (720° rotation in the clockwise direction) without including the epididymis. After torsion (2 h) and detorsion (4 h) periods, rats were sacrificed and bilateral orchidectomy was performed. Testis tissues were washed with cold saline solution, cut into small pieces with scissors, placed into glass bottles and homogenised in four volumes of ice-cold Tris-HCl buffer. Clear supernatant fluid was used for biochemical analyses. Treating rats with CAPE (applied at 10 µmol/kg, 30 min prior to T/D) attenuated the testicular injury, as well as the increase in the tissue levels of myeloperoxidase and thiobarbituric acid-reactant substances (TBARS) caused by T/D in the testis. Testis tissues showed a significant increase in glutathione peroxidase (GSH-Px) activity compared to the torsion group when CAPE was applied. Taken together, our results clearly demonstrate that CAPE treatment exerts a protective effect on testicular T/D, and part of this effect may be due to inhibiting the neutrophil-mediated cellular injury.