Changes in the expression of claudins and transepithelial electrical resistance of mouse Sertoli cells by Leydig cell coculture

In the testis, tight junctions (TJs) between adjacent Sertoli cells are important for the formation of blood-testis barrier (BTB). To verify the role of paracrine interactions between the Sertoli and Leydig cells in the structure and function of BTB in testis, the expression of claudin-1 and -11, and transepithelial electrical resistance (TER) of the mouse Sertoli cells were examined under the Leydig cell coculture. TER of Sertoli cell monolayer was significantly larger under the Leydig cell coculture in comparison with the control culture. Meanwhile, the expression of claudin-1 slightly decreased and claudin-11 significantly increased in the Sertoli cells in the Leydig cell coculture compared with control. Testosterone significantly increased claudin-11 expression in cultured Sertoli cells. Taken together, it suggested that Leydig cell coculture changed the structure and functions of inter-Sertoli TJs in vitro. Interactions between Leydig and Sertoli cells might be involved in the development of functional blood testis barrier in mouse testis.     

Microscopic and Ultrastructural Changes of the Intestine in Abdominal Compartment Syndrome

Background: The purpose of the study was to examine the microscopic changes of intestine villus and ultrastructural changes of tight junction in the presence of intra-abdominal hypertension (IAH) of 20 mmHg. Materials and Methods: Forty Sprague-Daley rats (250 ± 25 g) divided into four groups (n = 10) were studied. In 30 animals, the intra-abdominal pressure was increased to 20 mmHg for 1 hr, 2 hr, and 4 hr, respectively. The others served as controls. The specimens of the intestines were submitted to histopathology using hematoxylin and eosin staining, and to ultrastructural analysis using a scanning electron microscope. Results: The histopathological study revealed different degrees of changes ranging from swelling and degeneration of villous epithelial cells to extensive denudation and collapse of the villus. Ultrastructural analysis revealed dilatation of the tight junctions and paracellular spaces in the presence of IAH. The severity of the damage was related to the time of IAH. Conclusions: Increased IAH caused a significant damage to the intestinal epithelium and a marked dilatation of intestinal tight junction, leading to the increased mucosal barrier permeability. It may explain why IAH was often associated with bacterial translocation and sepsis.     

紧密连接蛋白2缺失对小鼠胆汁及胆固醇结石形成的影响分析

目的 旨在探究紧密连接蛋白2（Cldn2）在胆汁分泌中的相关作用。方法 实验将受试小鼠分为Cldn2^-/-基因敲除组（Cldn2^-/-组）与Cldn2^+/+野生型组（Cldn2^+/+组）,进行4周致石饮食,两组小鼠分别获得肝脏及胆管组织,通过组织学、生物化学、电生理学分析等分析两组小鼠肝脏和胆管内水及相关离子渗透压梯度运动差异;评价两组小鼠胆结石的形成情况。结果 Cldn2 ^+/+组小鼠的胆汁流速为[（58.0±4.4）μL/ （min·kg）],约为Cldn2^-/-组[29.0±3.3 μL/（min·kg）]的2倍,Cldn2^-/-组胆汁中总胆固醇、磷脂、总胆汁酸及总胆红素均显著高于Cldn2+/+组（P＜0.01）。当将Cldn2^+/+组的肝内胆管单位（IBDU）置于低渗缓冲液中时,腔内空间立即增加且持续至90 s;相反,当IBDU置于高渗缓冲液中时,Cldn2^+/+组腔内空间立即降低并且持续收缩至90 s;Cldn2^-/-组具有相同的趋势,但变化幅度显著低于Cldn2^+/+组,表现为90 s低渗时[（1.20±0.02）% vs（1.34±0.02）%,P=0.035],高渗时[（0.67±0.01）% vs（ 0.82±0.01）%,P=0.025]。与Cldn2^+/+组比,Cldn2^-/-组Na⁺浓度降低,但K⁺浓度升高,跨上皮电传导速度显著降低。致石饮食后所有Cldn2^-/-组肉眼均可见明显的胆结石形成,结石的主要成分是胆固醇（＞98%）,而Cldn2^+/+组中仅1 只小鼠有结石形成。结论 Cldn2 可能通过调控胆汁中的水分、胆固醇含量以及离子浓度,进而影响胆汁流速和成分,该蛋白的缺失会导致胆固醇结石发生的风险增加。     

Is testosterone involved in the initiation of spermatogenesis in humans? A clinicopathological presentation and physiological considerations in four patients with Leydig cell tumours of the testis or secondary Leydig cell hyperplasia

The purpose of this study was to evaluate the role of testosterone on the puberal development of spermatogenesis and to present additional clinicopathological data which bring about new information to this controversial subject. Four pre-pubertal patients are presented, 2 of them bearing Leydig cell tumours of the testis in the form of nodular masses. In both cases seminiferous tubules in the immediate vecinity to the tumours showed complete development of spermatogenesis, while those located away from the tumours were infantile in nature. Gonadotrophic levels were within the normal pre-pubertal range in these 2 cases. In one of the patients, testosterone concentration in the testis showed higher values than normal, and a concentration gradient was detected between the tumoral nodule and non-tumoral parenchyma. The 3rd patient had a pineal choriocarcinoma producing high amounts of hCG and consequently a diffuse hyperplasia of Leydig cells with high levels of plasma testosterone. Seminiferous tubules showed development up to pachytene spermatocytes. The last case was a precocious puberty in a boy with a tumour of the 3rd ventricle area. He had elevated levels of testosterone in the testis and plasma. In the testicular biopsy, stimulation of Leydig cells was detected. The seminiferous tubules showed mature Sertoli cells and pachytene spermatocytes. FSH levels were abnormally low. These 4 cases present in common different situations in which abnormally high amounts of testos-happens in the immature rat, the interaction between testosterone and gonadotrophins is essential for the normal initiation of spermatogenesis in normal puberty. Considerations are discussed on the possible synergistic role of gonadotrophins or other factors in relation with stimulation of seminiferous tubules by testosterone.     

Correlation of the hypo-osmotic swelling test, semen score and the zona-free hamster egg penetration assay in humans

The correlation between the hypo-osmotic swelling test, the modified Eliasson score for human semen analysis and the hamster egg penetration assay was examined. The results showed a weak but significant correlation between the group of subjects with swelling rates below 50% and the above 80% group. These groups showed hamster egg penetration assay rates of 9 +/- 14% and 39 +/- 29%, respectively (P less than 0.035). A significant correlation was also found between modified Eliasson scores (17 +/- 10 and 5 +/- 7, respectively) and swelling rates below 50% and above 70% (P less than 0.001). These results suggest that spermatozoa showing a low swelling rate have a poor fertilizing capacity. Therefore, the swelling tests and analysis of the patients semen can be used to discriminate sperm quality.     

Developmental comparisons of murine secretory amelogenesis in vivo,as xenografts on the chick chorio-allantoic membrane,and in vitro

Summary The temporal, spatial, and cytological characteristics of secretory amelogenesis in developing mouse mandibular first molar tooth organs have been compared during in vivo odontogenesis (from the Cap Stage in Theiler stage 25 C57BL/6 embryos to 10-day-old postnatal mice), as xenografts on the chick chorioallantoic membrane (CAM) for periods up to 7 days, and as explants in chemically defined medium without serum or antibiotics for periods up to 21 days in vitro. Tooth morphogenesis and cytodifferentiation proceeded in each environmental condition in the same sequence albeit at different rates of development. In vivo and CAM xenografts were remarkably comparable in their respective expressions of dentinogenesis and amelogenesis, whereas those explants cultured in a chemically defined medium without serum or antibiotics developed at a much slower rate (e.g., 0.5 days in vivo is equivalent to 1 day in vitro). In each experimental group, secretory amelogenesis was typically first detected along the mesial-buccal cusp of the molar organ independent of which environment was evaluated. Tooth morphogenesis in vitro and as xenografts on the CAM was routinely smaller than in situ odontogenesis. In each environmental condition a “stippled” precursor ultrastructural form of enamel matrix preceded mineralization, except during in vitro cultures of tooth organs. In vitro secretory amelogenesis or dentinogenesis did not indicate morphological characteristics of mineralization; both dentine and enamel matrices did not mineralize under the permissive environmental conditions used in these experiments. Calcium hydroxyapatite crystal formations within dentine and along the dentinoenamel junction during initial enamel matrix formation were not observed during in vitro tooth organogenesis, even for periods up to 21 days in vitro. We conclude that cap stage mandibular first molar tooth organs (enamel organ epithelium and adjacent dental papilla mesenchyme) from Theiler stage 25 embryos contain all of the necessary developmental instructions to express morphogenesis and cytodifferentiation except cues for the serum-containing factors for mineralization.     

Differences in osmolality, pH, buffering capacity, superoxide dismutase and maintenance of sperm motility in human ejaculates according to the degree of coagulation

The purpose of this study was to compare various seminal plasma parameters in fresh human ejaculates exhibiting different amounts of coagulum. Poorly coagulating samples demonstrated significantly lower osmolality and buffering capacity but had a higher pH than did samples with good coagulation. However, no correlation was obtained between the activity of superoxide dismutase and the amount of coagulum. Eight hours after ejaculation, sperm motility had decreased by 15 and 80% in samples with good and poor coagulation, respectively. It is suggested that subfertility may be associated with poor coagulation of ejaculates.     

Sperm acrosome antigen-1, a molecule intimately involved in the regulation of the acrosome reaction: analysis of expression on spermatozoa from infertile couples

Summary.  Sperm acrosome antigen-1 (SAA-1) is a molecule on the acrosomal cap of sperm from the human and a number of mammalian and lower species. SAA-1 was initially characterized by a monoclonal antibody (mab) AG7 directed against SAA-1. Previous studies indicate that SAA-1 may play an important role in the regulation of the acrosome reaction in the human and other species. Unselected couples seeking infertility treatment were subjected to an analysis of the amount of SAA-1 present on washed husband sperm. Using indirect immunonuorescence as well as radioimmunobinding assay, the expression of SAA-1 on patient spermatozoa was found to be significantly decreased compared to a group of healthy sperm donors. The decrease in SAA-1 did not correlate well with sperm morphology. Couples entered into the study were followed for an average of 12 months, while they received infertility treatment. Most couples conceived after a variable number of treatment cycles. It is concluded that a decrease of SAA-1 expression may contribute to subfertility, which can be overcome by the aid of assisted reproduction.     

Vascularized pancreas transplantation in the rat - details of the microsurgical techniques, results, and complications

Abstract. A detailed survey of the microsurgical techniques of vascularized pancreas transplantation in streptozotocin-diabetic rats is presented, based on the experience of 1450 transplants. Various techniques were employed to manage exocrine graft secretion. These included enteric, urinary, and free intraperitoneal drainage, as well as duct ligation and obstruction. In addition to general complications, such as anesthetic deaths, afterbleeding, and infection, a number of specific surgical complications, including graft pancreatitis, acute venous thrombosis, and hypoglycemia, occurred. The overall success rate of pancreas transplantation was 83. 9%. Incidence, causes, and possible prophylaxis of early and late postoperative complications are discussed in detail.     

DOX新辅助化疗方案对进展期食管胃交界部癌和远端胃癌的效果评价

目的探讨DOX方案新辅助化疗在进展期食管胃交界部癌（adenocarcinomaofesopha—gogastricjunction,AEG）及远端胃癌（distalgastriccancer,DGC）治疗中的作用。方法收集2008年10月至2011年3月间确诊的进展期胃癌患者58例,其中食管胃交界部腺癌18例（AEG组）,远端胃癌40例（DGC组）,术前行DOX方案化疗2周期,于化疗后14—21d行手术治疗。结果两组患者均完成术前新辅助化疗,化疗后AEG组CR3例,PR12例,SD3例,PD0例,总有效率为83．3％;DGC组CR2例,PR20例,SD17例,PD1例,总有效率55．0％,两组差异有统计学意义（P〈0．05）。AEG及GC的R0切除率分别为88．8％及87．5％,差异无统计学意义（P〉0．05）。两组化疗主要不良反应为粒细胞减少、胃肠道不良反应及外周神经毒性;术后并发症发生率差异无统计学意义（均P〉0．05）。两组1年生存率均为100％;AEG组2年生存率（77．8％）比DGC组（75．0％）亦无统计学差异（均P〉0．05）。结论DOX方案新辅助化疗对食管胃交界部癌较远端胃癌效果更好,缩小肿瘤、降低分期更明显,但远期疗效尚待进一步随访。     

Inhibition of Glucose Catabolism in Rat, Hamster, Rhesus Monkey and Human Spermatozoa by α-Chlorohydrin

Spermatozoa from the cauda epididymidis of rats treated with RS -α-chlorohydrin (10 mg/kg/day p. o. × 7) had the same ATP content as control spermatozoa immediately after collection, but were unable to metabolize glucose in vitro and so their ATP content declined more rapidly than that of control spermatozoa.
The in vitro metabolism of glucose by spermatozoa from rat, hamster, rhesus monkey and human was inhibited by 80% or more after 15–30 min preincubation in the presence of RS-α-chlorohydrin concentrations of < 1, > 10, 5–10 and > 50 mM, respectively. Inhibition of glucose oxidation was correlated with a reduction in ATP concentration in the spermatozoa.
These data support the hypothesis that α-chlorohydrin acts as an antifer-tility agent by inhibition of sperm glycolysis but suggest that RS -α-chlorohydrin would not be an effective contraceptive in man. However, the glucose metabolism of human spermatozoa was significantly inhibited by < 10 mM of the S -enantiomer of α-chlorohydrin.     

Extracorporeal ("ex vivo") connection of pig kidneys to humans. III. Studies of plasma complement activation and complement deposition in the kidney tissue

ABSTRACT: The complement system is one of the important factors involved in the hyperacute rejection of xenografts. This report deals with the activation of the complement system in a clinical trial where pig kidneys were extracorporeally connected to two volunteer dialysis patients who were pretreated with plasmapheresis in order to substantially reduce anti-pig xenoantibodies. The clinical data of the perfusion experiments and the patients humoral immune response to pig xenoantigens have been reported in detail (Xenotransplantation 1996; 3:328–339, 340–353). Three consecutive daily plasmapheresis treatments of the patients reduced the plasma complement protein (C3, C4, and C5) concentrations to 8–27% of the baseline values. The perfusion of the pig kidney connected to patient 1 was terminated at 65 min due to graft rejection and this patient was not hemodynamically affected by the experiment. The second experiment was terminated at 15 min due to an anaphylactic like reaction of the patient. In patient 1 a slight reduction of plasma C3, C4, and C5 and an increase of C5a and SC5b–9 occurred, while C3a decreased during the perfusion. Patient 2 had an increase of all complement parameters, most prominent for C4d and SC5b–9, which occurred concomitant with the appearance of the anaphylactic like side effects. In general, plasma levels of PMN elastase, IL6 and IL8 increased in both patients during the perfusion. Immunohistochemical investigation of the kidney tissues revealed deposition of human complement factors C1q, C4c, and C3c in a congruent pattern with the vasculature of the kidney in patient 1. In kidney 2 only trace amounts of C1q and C3c were found. Both kidneys were negative for properdin. Therefore, in this experimental set up with extracorporeal connection of pig kidneys to the human circulation the human complement cascade is activated mainly through the classical pathway.     

Results of the section of the filum terminale in 20 patients with syringomyelia,scoliosis and Chiari malformation

Summary Background. Spinal cord traction caused by a tight filum terminale may be considered a pathogenic mechanism involved in the development of syringomyelia, the Chiari malformation (type I) and scoliosis. Section of the filum terminale is proposed as a useful surgical approach in these conditions.Methods. Between April 1993 and July 2003, a total of 20 patients (8 men and 12 women) with a mean age of 33.5 years underwent section of the filum terminale with or without opening of the dural sac through a standard sacrectomy. Eight patients suffered from scoliosis, 5 from syringomyelia, 2 from Chiari malformation and 5 with a combination of these conditions.Finding. After section of the filum terminale, patients with syringomyelia showed an early clinical improvement of dysaesthesia, thermo-anaesthesia, hypo-aesthesia and walking difficulties. Rising of the medullary conus was also observed. In patients with scoliosis, back pain improved dramatically and a curve reduction was noticed, although progression of the curve was observed in one case. In patients with Chiari malformation, headache, dysaesthesia and paraparesis disappeared.Conclusions. Section of the filum terminale is a useful strategy in the treatment of scoliosis, syringomyelia and the Chiari malformation, and offers a new aetiological basis for the understanding of these three disorders.     

BLOOD-TESTIS BARRIER: Maintained Function of Inter-Sertoli Cell Junctions in Experimental Cryptorchidism in the Rat, as Judged by a Simple Lanthanum-Immersion Technique

In order to elucidate the mechanisms behind the deleterious effects on the germinal epithelium of experimental cryptorchidism the hypothesis that a leaking blood-testis barrier is the cause of the damage was tested. The permeability of the specialized inter- Sertoli cell junctions to lanthanum after experimental cryptorchidism for 0.5 to 12 days was studied in the rat. In none of the time periods studied lanthanum had penetrated beyond the inter-Sertoli cell junctions. A simple lanthanum immersion technique was used. Testes of 15-days old rats (before the development of the barrier) were used as a positive control of the method, and in these testes lanthanum had penetrated up to the future lumen.

Zusammenfassung

Um den Mechanismus zu erklären, der für die schädigenden Einflüsse auf das Keimepithel bei experimentellem Kryptorchismus verantwortlich ist, wurde die Hypothese untersucht, wonach eine durchlässige Blut-Hoden-Schranke Ursache dieser Schäden ist. Die Permeabilität der besonders ausgebildeten Inter-Sertoli-Zell-Verbindungen für Lanthanum nach exp. Kryptorchismus wurde 0, 5 bis 12 Tage lang bei Ratten studiert. Zu keinem Zeitpunkt dieser Studie penetrierte Lanthanum durch die Inter-Sertoli-Zell- Verbindungen. Methodisch wurde eine einfache Lanthanum-Immersions-Technik benutzt. Man verwendete 15 Tage alte Ratten (vor der Entwicklung der genannten Schranke) als positive Kontrolle; in diesen Fällen war Lanthanum bis in das Lumen penetriert.     

Cytological and functional studies of preosteoclasts and osteoclasts in the alveolar bones from neonatal rats using microperoxidase as a tracer

Summary Microperoxidase (MP) was used to investigate the cytological and functional features of preosteoclasts and osteoclasts during rat alveolar bone development. We observed mononuclear cells as preosteoclasts and multinuclear cells with and without ruffled borders (RB). In the bone facing multinuclear cells with RB as active osteoclasts, MP was extensively deposited along the external bone matrix undergoing resorption, and was phagocytosed with bone components into the vacuoles of osteoclasts. Neither preosteoclasts nor multinuclear cells without RB took up MP and bone components. Only multinuclear cells with RB seemed to resorb bone. Monocytes/macrophages (MMP) phagocytosed MP through all regions of the plasma membrane, whereas osteoclasts took up MP only through the RB which was a part of the plasma membrane. Endogenous peroxidase was detected in the MMP but not in preosteoclasts and osteoclasts. Thus, osteoclasts were considerably different from the MMP. The numbers of MMP were extremely few close to the osteoclasts, whereas moderate numbers of preosteoclasts were located close to the osteoclasts. Except for the nucleus and RB, there were many morphological similarities between preosteoclasts and osteoclasts. We therefore suggest that preosteoclasts, rather than MMP, are the precursors of osteoclasts during alveolar bone development of neonatal rats.