SPAG5 promotes proliferation and suppresses apoptosis in bladder urothelial carcinoma by upregulating Wnt3 via activating the AKT/mTOR pathway and predicts poorer survival.

Sperm-associated antigen 5 (SPAG5) is involved in various biological processes. However, the roles of SPAG5 in bladder urothelial carcinoma (BUC) are unknown. This study showed that upregulation of SPAG5 was detected frequently in primary BUC tissues, and was associated with significantly worse survival among the 112 patients that underwent radical cystectomy (RC). Up and downregulating the expression of SPAG5 enhanced or inhibited, respectively, the proliferation of BUC cells in vitro and in vivo, and suppressed or enhanced, respectively, apoptosis in vitro and in vivo. Moreover, SPAG5 increased the resistance of BUC cells to chemotherapy-induced apoptosis. Mechanistic investigations showed that SPAG5 promotes proliferation and suppresses apoptosis in BUC at least partially via upregulating Wnt3 through activating the AKT/mTOR signaling pathway. The importance of the SPAG5/AKT-mTOR/Wnt3 axis identified in BUC cell models was confirmed via immunohistochemical analysis of a cohort of human BUC specimens that underwent RC. Collectively, our data suggested that in patients with BUC who underwent RC, high SPAG5 expression is associated with poor survival. In addition, targeting SPAG5 might represent a novel therapeutic strategy to improve the survival of patients with BUC.

[Binding and interaction of histones and transcription factors on the promoter of hAMFR gene].

In this study, histone H1, core histones H2A-H2B and H3-H4 were purified from chicken erythrocytes by hydroxylapatile chromatography. The nuclear extract was prepared from HeLa cells. We investigated the binding and interaction of histones and transcription factors on the upstream sequence of human autocrine motility factor receptor (hAMFR) gene by gel shift mobility assay. We found that the binding of H1 on the promoter sequence of hAMFR gene was relatively stable. We propose that H1 plays an important role in stablizing chromatosome. We also found that histones and HeLa cell extract could form a ternary complex with the DNA template.

[Effects of the interaction between histone and hAMFR gene promoter on the transcription activity in vitro].

The effects of interaction between histones and human autocrine motility factor receptor (hAMFR) gene promoter on the transcription activity in vitro was investigated by using histones purified from chicken erythrocytes, HeLa cell nuclear extracts and heat-treated supernatants of Xenopus eggs. The results showed that the competitive binding of histones and transcription factors at the promoter of hAMFR gene was very important to the transcription in vitro. If a pre-initiation complex was formed with HeLa cell nuclear extracts on the promoter prior to nucleosome assembly, it would prevent nucleosome-mediated transcription repression. When the nucleosome was assembled on the promoter in advance, the transcription activity could be repressed. When histones and HeLa cell nuclear extracts were mixed in the reaction simultaneously, the transcription activity would depend on the relative amount of histones to that of HeLa cell nuclear extracts.

Cell proliferation and oncogene expression after bile duct ligation in the rat: evidence of a specific growth effect on bile duct cells.

The proliferative response of the rat liver was measured after temporary or permanent total biliary obstruction (BDO) and in different regions after selective ligation of the lobar ducts draining the right 60% of the hepatic mass. The results were compared with those after 70% partial hepatectomy (PH). Cell proliferation was assessed globally by measuring DNA synthesis and stratified to the separate cell populations with cytostaining techniques that allowed distinction of hepatocytes, duct cells, and nonparenchymal cells (NPCs). In selected experimental groups, gene expression was determined of transforming growth factor-beta 1 (TGF beta-1), prothrombin, c-erb-B2, transforming growth factor alpha (TGF alpha), human Cyclophilin (CyP), and 28S ribosomal RNA. The stimulation of a proliferative response to total BDO required obstruction for longer than 24 hours, but after this deligation did not switch off regeneration. In the first week after permanent BDO, there was progressive infiltration of NPCs, fibrous linkage of some portal areas, and a crescendo of DNA synthesis that was obvious at 24 hours, maximal at 48 hours, and back nearly to baseline at 6 days. At the 2-day mark, the bile duct cells had a 17-fold increase in proliferation, accompanied by a threefold to fourfold increase in hepatocyte renewal. Little or no increase in expression of TGF alpha or the hepatocyte-specific prothrombin gene was detectable in the first 48 hours, whereas levels of the oncogene c-erb-B2 that is associated with cholangiocarcinoma were expressed from 48 to 96 hours.(ABSTRACT TRUNCATED AT 250 WORDS)

Modified simple cold storage of rat livers with UW solution.

Rat livers were preserved with the conventional use of UW solution for 30, 42, and 48 hr and compared with livers in which the vascular bed was expanded with an additional 10 to 60 ml UW/100 g liver. The extra UW, expressed as % liver weight, was entrapped during final portal infusion by typing off the supra- and infrahepatic inferior vena cava. A beneficial influence of the vascular expansion was most pronounced in the 40% group, with 10/10, 5/10, and 3/10 long-term survivors following transplantation after 30, 42, and 48 hr preservation versus 3/10 and 0/10 after 30 and 42 hr in the 0% controls. In separate experiments, surrogate indices of preservation quality following reperfusion explained this effect. The 40%--and, to a lesser extent, 20%--livers had higher and more uniformly distributed portal blood flow, better tissue oxygenation, smaller increases in postperfusion liver enzymes, higher adenine nucleotides and energy charge, and less histopathologic evidence of hemorrhage and congestion. Pressure changes in the vena cava fluid sump in additional experiments indicated that retrograde infusion of the trapped UW solution occurred in all of the 10-60% groups during the first 6 hr with stable pressures of 1.5 to 3 cm H2O thereafter. Collectively, these data suggest that the much discussed selective vulnerability of the microvasculature of stored allografts is due in part (or principally) to its selective lack of long-term exposure to the UW solution, which drains out of the open vessels but not from the parenchyma. The potential clinical exploitation of this concept is discussed.

Screening for candidate hepatic growth factors by selective portal infusion after canine Eck's fistula.

Completely diverting portacaval shunt (Eck's fistula) in dogs causes hepatocyte atrophy, disruption of hepatocyte organelles, fatty infiltration and low-grade hyperplasia. The effect of hepatic growth regulatory substances on these changes was assessed by constantly infusing test substances for four postoperative days after Eck's fistula into the detached left protal vein above the shunt. The directly infused left lobes were compared histopathologically with the untreated right lobes. In what has been called an hepatotrophic effect, stimulatory substances prevented the atrophy and increased hepatocyte mitoses. Of the hormones tested, only insulin was strongly hepatotrophic; T3 had a minor effect, and glucagon, prolactin, angiotensin II, vasopressin, norepinephrine and estradiol were inert. Insulin-like growth factor, hepatic stimulatory substance, transforming growth factor-alpha and hepatocyte growth factor (also known as hematopoietin A) were powerfully hepatotrophic, but epidermal growth factor had a barely discernible effect. Transforming growth factor-beta was inhibitory, but tamoxifen, interleukin-1 and interleukin-2 had no effect. The hepatotrophic action of insulin was not altered when the insulin infusate was mixed with transforming growth factor-beta or tamoxifen. These experiments show the importance of in vivo in addition to in vitro testing of putative growth control factors. They illustrate how Eck's fistula model can be used to screen for such substances and possibly to help delineate their mechanisms of action.

Orthotopic liver transplantation and the cytosolic estrogen-androgen receptor status of the liver: the influence of the sex of the donor.

Mammalian liver is known to contain cytosolic receptors for both estrogens and androgens. Furthermore, certain mammalian hepatic functions are known to display a sexual dimorphism. However, in clinical liver transplantation, the sex of the donor is not taken into consideration in selection of the donor. In this study, the effect of liver transplantation on the estrogen and androgen receptor content of the liver was determined. Adult male and female rats were subjected to orthotopic liver transplantation, using donors from both the same and the opposite sex as the recipient. The animals were killed on the tenth postoperative day, and the livers were assayed to determine their cytosolic estrogen and androgen receptor content. Transplantation of a liver from a male donor into a male recipient, from a male donor into female recipient and from a female donor into a male recipient produced similar changes in the number of cytosolic estrogen and androgen receptors in hepatic cytosol. In all three situations, the estrogen receptor content in the cytosol of the transplanted liver was the same as that found in an unoperated male liver, and the cytosolic content of the androgen receptor was the same as that of an unoperated female liver. After transplantation of the liver from a female donor into a female recipient, the estrogen and androgen receptor content in the cytosol of the transplanted liver was the same as that of an unoperated female.(ABSTRACT TRUNCATED AT 250 WORDS)

Cholesterol synthesis in bypassed segments of the small intestine in hyperphagic rats.

Previous studies have demonstrated that a variety of conditions that result in an increase in food intake lead to an increase in small-intestinal cholesterol synthesis. In the present study, it was determined whether hyperphagia induces an increase in cholesterol synthesis in segments of the small intestine excluded from contact with the food stream and whether this increase would occur in bypassed segments of the proximal or mid-small intestine. In hyperphagic diabetic rats, cholesterol synthesis is increased 91% in the proximal portion of the small intestine excluded from contact with nutrients. In lactating rats, another model of hyperphagia, cholesterol synthesis is increased 2.4-fold in midintestinal segments excluded from contact with the food stream and 2.9-fold in segments of the proximal intestine that have been bypassed. These observations demonstrate that the hyperphagia-induced increase in small-intestinal cholesterol synthesis will occur in portions of the small intestine, even if contact with the food stream is prevented. In addition, this data demonstrated that the mass of the bypassed portion of the small intestine is increased in hyperphagic animals. In diabetic animals, the weight of the bypassed proximal intestine is increased 2.1-fold, whereas in lactating animals the mass is increased 50% in the bypassed midintestine and 74% in the bypassed proximal small intestine. In conclusion, the present study suggests that circulating or neurologic factors, or both, play a role in stimulating intestinal cholesterol synthesis in hyperphagic animals. These findings also suggest that indirect factors play a role in the increase in intestinal mass associated with hyperphagia.

Thyroid cancer: the case for total thyroidectomy.

Since there are no prospective studies concerning the treatment of thyroid cancer, there continues to be a considerable disagreement about the 'best' or most appropriate form of surgical treatment for patients with papillary or follicular thyroid cancer. Some surgeons recommend selective treatment depending upon the type of thyroid tumor and stage of the disease. Some advocate thyroid lobectomy and isthmusectomy, some near total thyroidectomy, and some total thyroidectomy for patients with papillary and follicular thyroid cancer. Total thyroidectomy for thyroid cancer would be the treatment of choice for virtually all patients with thyroid cancers if it could be done without complications. We therefore reviewed 160 consecutive patients who had total thyroidectomy for suspected or proven thyroid cancer to determine the complication rate of total thyroidectomy. One hundred and three patients had primary operations, 57 had reoperations with completion of total thyroidectomy and 124 had thyroid cancer. Serious complications (i.e. vocal cord paralysis or hypoparathyroidism) included two cases of transient bilateral recurrent nerve palsy, two patients with presumed transient unilateral vocal cord paralysis, three recurrent laryngeal nerves that were purposely sacrificed because of invasion of the nerve, and one case of permanent hypoparathyroidism. Two other patients developed postoperative wound infections. Only one of the permanent complications, the case of permanent hypoparathyroidism, could have been avoided by a lesser procedure. The experienced surgeon can perform a total thyroidectomy with minimal morbidity, and this procedure has certain theoretical and practical advantages. It should not be done, however, if it will result in a significant complication rate and, in selected patients, it may be preferable to leave a small amount of thyroid tissue to protect the blood supply to the parathyroid glands or recurrent laryngeal nerve.

Localizing studies in patients with persistent or recurrent hyperparathyroidism.

Preoperative localizing studies are essential for patients with persistent or recurrent hyperparathyroidism requiring reoperation, because of loss of normal tissue planes and because the hyperfunctioning parathyroid tissue that remains is more likely to be situated in an ectopic position. The value of noninvasive and invasive localizing techniques was evaluated in 59 consecutive patients undergoing reoperation for persistent (40 patients) or recurrent (19 patients) hyperparathyroidism. Magnetic resonance imaging was performed in 17 patients; 11 results (65%) were positive, 3 (18%) were negative, and 3 (18%) were false-positive. Ultrasonography was performed in 52 patients; 29 (56%) were positive, 16 (31%) were negative, and 7 (13%) were false-positive. Computed tomography was performed on 41 patients; 19 (46%) were positive, 16 (39%) were negative, and 6 (15%) were false-positive. Thallium chloride 201-technetium 99m pertechnetate scans were used in 39 patients; 19 (49%) were positive, 11 (28%) were negative, and 9 (13%) were false-positive. One or more of these noninvasive tests was positive in 78% of the cases. Highly selective venous catheterization with measurement of immunoreactive parathyroid hormone concentration localized the abnormal parathyroid gland in 20 of 28 patients (71%) overall and in 8 of the 14 patients (57%) whose tumors were not identified by the noninvasive techniques. Since false-positive results were common, a combination of localizing studies was helpful in identifying the abnormal gland. Fifty-three of the 59 patients (90%) were successfully treated at the initial reoperation and three were successfully treated at a second reoperation. Advances in parathyroid localization have contributed to the improved surgical results in patients with persistent or recurrent hyperparathyroidism.