Objective To establish a new model on isolated human cadaver testes with ischemiareperfusion (I/R). MethodsThirteen isolated cadaver testes contributed by 13 persons were preserved under 0℃-4℃ hypothermia and then reperfused under 37℃. Histological and histochemical changes were observed. Results4℃ cold ischemia in 12 hours induced only trivial swelling and vascular degeneration of endothelial cells (ECs), obvious pathologic changes occurred after 24 hours, including detachment of ECs, separation between basement membrane and seminiferous epithelium, degeneration and detachment of spermatogenous cell and edema of mesenchyme. Injury was worse along with the prolongation of cold preservation time. Changes of LDH and SDH activities were found by histochemical staining. Reperfusion following 6 hours ischemia induced tissue injury and unusual enzyme activity. All changes were more obvious after reperfusion following 12,18,24 or 36 hours cold ischemia.Conclusion This new model on isolated cadaver testes with ischemiareperfusion is successful, it can substitute other solid organs of human beings for I/R injury study.
To explore the effect of burying testis in inguinal pocket on spermatogenesis. Methods Sixty New Zealand rabbits of 6-8 months old included 36 males and 24 females, weighing 2.5-2.7 kg. The male rabbits were randomly divided into the experimental group (n=18)and the control group (n=18). The model of repairing skin defect of scrotum were establ ished by burying testes in inguinal region subcutaneously in the experimental group. The rabbits were not treated in the control group. The sperms were collected and the surface temperature of testis was measured in both groups after 8 weeks. Testes biopsies were harvested from 6 rabbits of 2 groups randomly respectively. The apoptosis of spermatogeniccells was detected with TUNEL. The other 12 male rabbits in two groups were fed respectively with female rabbits to observe the fertil ity. Results The semen density and the spermid activity ratio were (237.3 ± 39.7) × 109/L and 76.9% ± 3.8% in the control group, and were (4.7 ± 2.7) × 109/L and 0 in the experimental group respectively; showing statistically significant difference between two groups (P lt; 0.05). The average superficial temperature of testes was (38.02 ± 0.36)℃ in the experimental group and (36.15 ± 0.64) in the control group (P lt; 0.05). TUNEL results showed: The spermatogenic epithel ium became thin and obvious apoptotic spermatogenic cells were found in experimental group; the spermatogenic epithel ium was normal and few apoptotic spermatogenic cells were found in the control group. The apoptotic index (AI) was 89.69% ± 3.76% in the experimental group and 7.73% ± 4.95% in the control group (P lt; 0.05). The Pairing results showed that the female rabbits pairing with male rabbits of the experimental group were all not pregnant, and those of the control group were all pregnant (P lt; 0.05). Conclusion As the same as the scrotum was reconstructed with skin flaps, it will induce the rabbit infertil ity that the testes were buried in inguinal region subcutaneously to repair defect of scrotum skin. The main reason is the excessive apoptosis of spermatogenic cell by the high testes environmental temperature.
Objective To investigate the feasibility and characteristic of tissue engineered testicular prosthesis with highdensity polyethylene(HDPE，trade name: Medpor) and polyglycolic acid(PGA). Methods The chondrocytes were isolated from the swine articular.The PGA scaffold was incorporated with medpor which semidiameters were 6mmand 4mm respectively.Then, the chondrocytes (5×10 7/ml) were seeded onto Medpor-PGA scaffold and cultured for 2 weeks. The ten BALB／C mice were divided into two groups randomly(n=5). In the experimental group, the cell-scaffold construct was implanted into subcutaneous pockets on the back of nude mice. In the control group, the Medpor-PGA scaffold was implanted. The mice of two groups were sacrificed to harvest the newly formed cartilage prosthesis after 8 weeks. Macroscopy, histology and immunohistochemistry observations were made. Results The gross observation showed that on changes were in shape and at size, the color and elasticity were similar to that of normal cartilage and that the cartilage integrated with Medpor in the experimental group; no cartilage formed and fiberlike tissue was found in the control group. HE staining showed that many mature cartilage lacuna formed without blood vessel and some PGA did not degradated completely. Toluidine blue staining showed extracellular matrix had metachromia. Safranin O-fast green staining showed that many proteoglycan deposited and collagen type Ⅱ expression was bly positive. In the control group, Medpor was encapsulated by fiber tissue with rich blood vessel. Conclusion The newly formed complex of Medpor-PGA and cells was very similar to testicle in gross view and to normal cartilage in histology. This pilot technique of creating testicular prosthesis by incorporating tissue-engineered cartilage with Medpor demonstrated success.
Objective To establish dog model of testicular autotransplantation with a modified technique.Methods Testicular autotransplantations were performed on the right side of 30 male dogs, whose ages ranged from 1.5 to 2.0 years old and weights ranged from 14 to 17 kg. After the spermatic artery with a cuff of abdominal aorta and spermatic vein and with a cuff of inferior vena cava were detached, the testis was perfused and kept at icing temperature. An end-to-side anastomosis of the spermatic vessels to the external iliac vessels was conducted subsequently. The survival conditions of the auografts were assessed by digital subtraction arteriography (DSA). Histological examination and detection on the serum levels of follicle stimulating hormone (FSH), luteotrophic hormone (LH), and testosterone (T) were made at two weeks intervals. Results Of the 30 testicular autotransplantations performed, 27 cases were successful. The success rate was 90%. The time of heat ischemia, cold ischemia, anastomosis of spermatic vessels, and total operation was 4.5±0.9 minutes, 50.0±10.0 minutes, 35.5±5.5 minutes, and 3.5±0.5 hours respectively. DSA proved that the testis survived well. No morphological abnormality was found at different stages of the spermatogenic cells. The LH level was higherthan that before operation, being statistically different (Plt;0.05);however, the levels of FSH and T did not changed significantly (Pgt;0.05). Conclusion A stable and feasible model of testicular autotransplantation is established and it provides a reliable experimental platform for human testicular transplantation.
OBJECTIVE: To investigate the injury on isolated testes induced by ischemia/reperfusion(I/R), and the protective effect of Yisheng injection on the injury. METHODS: Twenty-six isolated cadaver testes contributed by 13 persons were preserved with 4 degrees C 250 ml hypertonic citrate alloxuric (HCA) solution and then reperfused with 37 degrees C 500 ml HCA. Solution of experimental group contained 500 micrograms/ml Yisheng injection. In simple cold preservation test, involving in 8 experimental and 8 control testes, a series of time points (6, 12, 18, 24, 36, 48, 60, 72 hours) were set to harvest. 10 testes (1 testis respectively on 6, 12, 18, 24 and 36 hours in experimental and control groups) were reperfused with 37 degrees C HCA for 6 and 12 hours. Histological and histochemical changes were observed. RESULTS: In the experimental testes, 4 degrees C cold preservation in 24 hours could not induce obvious pathologic changes. After 24 hours, changes such as swelling, vacuolar degeneration or detachment of endothelial cells (ECs), separation between basement membrane and seminiferous epithelium, mal-alignment of spermatogenous cell and edema of mesenchyme could be observed. In the testes preserved for 12 hours, the activity of lactic dehydrogenase(LDH) and succinic dehydrogenase (SDH) increased, then fallen after 24 hours. The activity of Nitric oxide synthetase(NOS) decreased after 18 hours. All changes were more obvious after following 37 degrees C reperfusion. In the control testes, swelling and vacuolar degeneration of ECs occurred on 12 hours cold preservation, and injury was worse along with the prolongation of cold preservation time. Pathologic changes of ECs, seminiferous epithelium and mesenchyme were serious after 37 degrees C reperfusion. CONCLUSION: 4 degrees C cold preservation in 24 hours can only cause mild ECs’ injury, and obvious abnormal testes’ histological profile can be observed beyond 24 hours. 37 degrees C reperfusion will make injury worse. Yisheng injection can keep isolated testes histologic structure well in 24 hours cold preservation, and it has protective effect on I/R injury.
From October 1985 to October 1988, fourty-nine impalpable testis in 39 patients were treated. The ages of the patients ranged from lyear 10 months to 11 years. There were fixed testis into scrotum (in 24 cases), superficial layer of aponeurosis of musculus obliquus externus abdominis (in 4 cases), autotransplantation (in 7 cases), and testectomy (in 11 cases). No testis in 3 cases were conformed in exploration. The author suggestes it necessary to do exploration early in impalpable testis of children to obtain good physiologic function of testis.
A clinical experience was introduced in reconstruction of the urinary bladder for 16 cases of tumor patients by pedieled testicular sheath. A satisfactory results were achieved in patients after a follow-up of 8 months to 5.5 years. The advantages were that: (1) the testicular sheath was an autogenous biomemhrane, it had no immunologic reaction; (2) rapid healing was achieved by vascularized testicular sheath; (3) the testicular sheath had a good elasticity; (4) there was no postoperative electrolytes disturbance because of the poor absorption of the sheath; (5)a rapid recovery of the bladdor function; and (6)patient felt normal when urination.
目的 通过检测人睾丸生殖细胞肿瘤中的Skp2蛋白质异常表达,探讨相关意义。 方法 应用S-P免疫组织化学法检测睾丸生殖细胞肿瘤，正常睾丸组织和慢性睾丸炎组织中Skp2的表达。 结果 睾丸生殖细胞肿瘤中Skp2阳性表达率为74.5%，正常睾丸组织中Skp2阳性表达率为20.0%，在慢性睾丸炎组织中Skp2阳性表达率为40.0%，在3种不同睾丸组织中表达差异有统计学意义（P＜0.05）；Skp2表达与不同组织学类型的睾丸生殖细胞肿瘤无相关性（P＞0.05）；随着临床分期的增高，睾丸生殖细胞肿瘤中的Skp2表达增多，差异无统计学意义（P＞0.05）。 结论 在人睾丸生殖细胞肿瘤中的Skp2高表达，提示细胞周期的异常调控在睾丸生殖细胞肿瘤的发生、分化中起着重要的作用。