{"title":"Expression of follistatin messenger ribonucleic acid in Sertoli cell-enriched cultures: regulation by epidermal growth factor and protein kinase C-dependent pathway but not by follicle-stimulating hormone and protein kinase A-dependent pathway.","authors":"U Michel, J Esselmann, E Nieschlag","doi":"10.1530/acta.0.1290525","DOIUrl":"https://doi.org/10.1530/acta.0.1290525","url":null,"abstract":"<p><p>In this study the localization and regulation of steady-state follistatin messenger ribonucleic acid (mRNA) levels in testicular cell cultures were examined with a solution-hybridization assay using a specific 32P-labelled cytosolic RNA antisense probe for follistatin and a 35S-labelled cytosolic RNA antisense probe for cyclophilin as internal standard. Testes from immature rats were dispersed with collagenase and fractionated in Sertoli and Leydig cell-enriched cultures. Follistatin mRNA was mainly localized to the Sertoli cell-enriched fraction and the expression of follistatin mRNA could be stimulated in vitro with fetal calf serum, epidermal growth factor or phorbol-12-myristate-13-acetate (an activator of protein kinase C), whereas follicle-stimulating hormone and forskolin (an activator of protein kinase A) had no effect. Neither prostaglandin E2, the synthetic glucocorticoid RU 28362 or all-trans-retinoic acid, which all regulate follistatin mRNA levels in non-testicular cell types, nor extracellular adenosine triphosphate (a purinergic receptor agonist) or testosterone had any obvious influence on follistatin mRNA levels in Sertoli cell-enriched cultures. From this study it is concluded that Sertoli cells are likely to be the source of follistatin expression in the rat testis, that follistatin mRNA levels in Sertoli cell-enriched cultures are subjected to regulation by epidermal growth factor and the protein kinase C-dependent pathway but are not regulated by extracellular adenosine triphosphate, follicle-stimulating hormone, all-trans-retinoic acid, prostaglandin E2, forskolin, testosterone or the glucocorticoid RU 28362 and that the regulation of follistatin mRNA is sex- and tissue-specific.</p>","PeriodicalId":6910,"journal":{"name":"Acta endocrinologica","volume":"129 6","pages":"525-31"},"PeriodicalIF":0.0,"publicationDate":"1993-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1530/acta.0.1290525","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"19101468","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
T P Links, J F Monkelbaan, R P Dullaart, T W van Haeften
{"title":"Growth hormone-, alpha-subunit and thyrotrophin-cosecreting pituitary adenoma in familial setting of pituitary tumour.","authors":"T P Links, J F Monkelbaan, R P Dullaart, T W van Haeften","doi":"10.1530/acta.0.1290516","DOIUrl":"https://doi.org/10.1530/acta.0.1290516","url":null,"abstract":"<p><p>A patient with acromegaly and hyperthyroidism due to a growth hormone-, thyrotrophin- and alpha-subunit-secreting pituitary adenoma is described. His decreased father had suffered from a pituitary tumour, and was likely to have had acromegaly as well. Plasma growth hormone and insulin-like growth factor I concentrations were elevated, with levels between 10 and 20 micrograms/l and 4.4 and 7.3 kU/l, respectively. In spite of hyperthyroidism (free thyroxine, 45 pmol/l; free triiodothyronine, 24 pmol/l), plasma thyrotrophin remained at 2.8 mU/l without any response to thyrotrophin-releasing hormone and could not be suppressed with exogenous administration of triiodothyronine. Plasma alpha-subunits were raised to 3.3-3.7 U/l (normal 0.4-1.1 U/l). Pathological examination of the surgically removed tumour showed a pituitary adenoma with the immunohistochemical presence of growth hormone, thyrotrophin, prolactin and alpha-subunit. This is the first report of a growth hormone-, thyrotrophin- and alpha-subunit-producing pituitary adenoma, which occurred in a familial setting.</p>","PeriodicalId":6910,"journal":{"name":"Acta endocrinologica","volume":"129 6","pages":"516-8"},"PeriodicalIF":0.0,"publicationDate":"1993-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1530/acta.0.1290516","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"19101466","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"Aromatase activity in the mare ovary during estrous cycle. Measurement of endogenous steroids and of their in vitro inhibitory effect.","authors":"H Amri, P Silberzahn, I al-Timimi, J L Gaillard","doi":"10.1530/acta.0.1290536","DOIUrl":"https://doi.org/10.1530/acta.0.1290536","url":null,"abstract":"<p><p>This present study was undertaken to clarify estrogen synthesis in the mare ovary. First of all, an evaluation of endogenous steroid contents was carried out in the follicular fluid and in the luteal tissue at different stages of the luteal phase. Radioimmunoassays were performed after separation and purification of each hormone by chromatography. High amounts of conjugated (0.9 mg/l) and unconjugated (4 mg/l) estradiol-17 beta were found in the follicular fluid of the large follicules (50 mm). These concentrations of estrogens decreased drastically in the luteal tissue, and only low levels of circulating estrogens are found during the luteal phase. On the other hand, a high aromatization ability has been evidenced in the cyclic corpus luteum in vitro. In an attempt to clarify the regulation of estrogen synthesis, we have tested the inhibitory effect of several endogenous steroids on equine ovarian aromatase activity. 5 alpha-Dihydrotestosterone appeared to be the most potent competitive inhibitor (Ki = 181 nmol/l) of aromatase activity, while the addition of a 3-sulfate group induced a slump in the inhibitory potency of estrone (Ki = 397 nmol/l vs 2206 nmol/l) and dehydroepiandrosterone (Ki = 291 nmol/l vs 6157 nmol/l). The physiological role of these conjugated steroids has not been known until now; we suggest that they would play a role in protecting aromatase from inhibition, in vivo. The high amounts of progesterone found in the luteal tissue (1.3 g/kg of proteins) might play a role in the regulation of estrogen production either by suppressing the induction of aromatase synthesis or by inhibiting the activity of the enzyme complex.</p>","PeriodicalId":6910,"journal":{"name":"Acta endocrinologica","volume":"129 6","pages":"536-42"},"PeriodicalIF":0.0,"publicationDate":"1993-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1530/acta.0.1290536","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"19101470","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"Comparison between the human serum growth hormone-binding protein and the water-soluble growth hormone-binding site released from IM-9 lymphocytes.","authors":"G Massa, M Ilondo, M Vanderschueren-Lodeweyckx","doi":"10.1530/acta.0.1290559","DOIUrl":"https://doi.org/10.1530/acta.0.1290559","url":null,"abstract":"<p><p>The characteristics of the human serum growth hormone-binding protein (GHBP) were compared with those of a water-soluble GH-binding site prepared by incubating cultured IM-9 lymphocytes in assay buffer with 25 mmol/l iodoacetamide. High-performance liquid chromatography gel filtration of the water-soluble GH-binding site incubated with 125I-labeled human GH ([125I]hGH) revealed a large peak of bound [125I]hGH eluting at the same position as the peak of [125I]hGH bound to the GHBP in serum. The estimated M(r) of the peak was 120,000, presumably representing one [125I]hGH bound to two binding sites. The binding specificities of the serum GHBP, the water-soluble GH-binding site and the GH receptor on IM-9 lymphocytes were identical. The binding affinities for 22,000 hGH and for 20,000 hGH of the serum GHBP were similar to the binding affinity of the water-soluble GH-binding site but lower than those of the cellular GH receptor. These findings show that the characteristics of the serum GHBP are comparable to those of the water-soluble GH-binding site released from IM-9 cells and support the hypothesis that in man the serum GHBP is produced by proteolytic cleavage of the cellular GH receptor.</p>","PeriodicalId":6910,"journal":{"name":"Acta endocrinologica","volume":"129 6","pages":"559-64"},"PeriodicalIF":0.0,"publicationDate":"1993-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1530/acta.0.1290559","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"19101473","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
F Azizi, A Sarshar, M Nafarabadi, A Ghazi, M Kimiagar, S Noohi, N Rahbar, A Bahrami, S Kalantari
{"title":"Impairment of neuromotor and cognitive development in iodine-deficient schoolchildren with normal physical growth.","authors":"F Azizi, A Sarshar, M Nafarabadi, A Ghazi, M Kimiagar, S Noohi, N Rahbar, A Bahrami, S Kalantari","doi":"10.1530/acta.0.1290501","DOIUrl":"https://doi.org/10.1530/acta.0.1290501","url":null,"abstract":"<p><p>In order to detect somatic and psychomotor disturbances in children and adolescents residing in areas of iodine deficiency, schoolchildren from three areas with different degrees of iodine deficiency were studied. In Randan, the prevalence of severe endemic goiter was accompanied by alteration in thyroid function, increased thyrotropin levels and retardation of both bone and psychomotor age and decreased intellectual quotient. In Tehran, where iodine deficiency is mild, visible goiter was present in 15% of schoolchildren but no alterations in thyroid function, serum thyrotropin, somatic or psychomotor development could be detected. In Zagoon, where the prevalence and severity of goiter was less than Randan but more than Tehran, thyroid function was normal but slightly decreased as compared to Tehran; somatic development was unaltered, but retardation in psychomotor development was evident and the mean intellectual quotient was less than that of Tehranian schoolchildren. These findings indicate the occurrence of physical and psychomotor disturbances in apparently normal schoolchildren from areas of iodine deficiency. Alteration in psychomotor development may occur in children with normal physical growth, due to iodine deficiency.</p>","PeriodicalId":6910,"journal":{"name":"Acta endocrinologica","volume":"129 6","pages":"501-4"},"PeriodicalIF":0.0,"publicationDate":"1993-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1530/acta.0.1290501","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"19100765","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
B Nygaard, P Gideon, H Dige-Petersen, N Jespersen, K Sølling, A Veje
{"title":"Thyroid volume and morphology and urinary iodine excretion in a Danish municipality.","authors":"B Nygaard, P Gideon, H Dige-Petersen, N Jespersen, K Sølling, A Veje","doi":"10.1530/acta.0.1290505","DOIUrl":"https://doi.org/10.1530/acta.0.1290505","url":null,"abstract":"<p><p>In order to throw light upon the eventual need for iodine supplementation in Denmark, four age groups of women (15, 30, 45 and 60 years) from the Holbaek municipality were invited for a clinical and ultrasound study of thyroid volume, structure and function. Of the 570 women invited, 391 accepted and were divided into the following groups: group I: 15 years, N = 113; group II: 30 years, N = 100; group III: 45 years, N = 98; group IV: 60 years, N = 80. The results were as follows the thyroid gland was palpable in 39% and visible in 16% of the entire group; 19% had a family history of thyroid disorders and 7.6% had a previous thyroid disorder. Thyroid volumes (median (range)) as measured by ultrasound were 12 ml (4-29 ml), 18 ml (5-47 ml), 18 ml (7-64 ml) and 18 ml (9-51 ml) in groups I-IV, respectively. The calculated 24-h iodine excretion was 65 micrograms (19-365 micrograms), 88 micrograms (15-274 micrograms), 97 micrograms (40-737 micrograms) and 83 micrograms (50-999 micrograms) in groups I-IV, respectively. An abnormal echo structure was present in 3, 10, 21 and 30%, respectively. Defining a goitre as a thyroid volume above 28 ml indicated a goitre prevalence of 17% in females aged 30-60 years in the Holbaek area of Denmark. Among the 60-year-old women, 3% had a clinically significant goitre (WHO grade III). Thyroid volume did not correlate with iodine excretion. The benefit of iodine supplementation is discussed.</p>","PeriodicalId":6910,"journal":{"name":"Acta endocrinologica","volume":"129 6","pages":"505-10"},"PeriodicalIF":0.0,"publicationDate":"1993-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1530/acta.0.1290505","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"19100766","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
H Ishida, K Suzuki, Y Someya, M Nishimura, C Sugimoto, M Goto, Y Taguchi, H Kasahara, S Kadowaki, H Imura
{"title":"Possible compensatory role of parathyroid hormone-related peptide on maintenance of calcium homeostasis in patients with non-insulin-dependent diabetes mellitus.","authors":"H Ishida, K Suzuki, Y Someya, M Nishimura, C Sugimoto, M Goto, Y Taguchi, H Kasahara, S Kadowaki, H Imura","doi":"10.1530/acta.0.1290519","DOIUrl":"https://doi.org/10.1530/acta.0.1290519","url":null,"abstract":"<p><p>Recent studies have revealed that altered mineral and vitamin D metabolism is observed in diabetic patients with the complication of osteopenia. In order to elucidate the role of parathyroid hormone-related peptide (PTHrP) on calcium homeostasis in diabetes, we have measured the serum level and urinary excretion of PTHrP as well as other serum calcium-regulating hormones in 106 patients with non-insulin-dependent diabetes mellitus (NIDDM) and 43 control subjects. The serum concentration of intact PTH was 2.34 +/- 0.13 (mean +/- SEM) pmol/l in NIDDM patients, which is significantly lower than the value of 3.11 +/- 0.14 pmol/l in the controls (p < 0.01). Both serum calcium and calcitonin, however, were not statistically different from controls. On the other hand, circulating PTHrP in NIDDM was 40.1 +/- 1.4 pmol/l, which is significantly elevated when compared to 27.3 +/- 1.3 pmol/l in the controls (p < 0.01). Moreover, urinary excretion of PTHrP also was significantly higher in NIDDM (p < 0.01). In the present study, the circulating calcium level was well preserved in NIDDM patients, although the PTH levels were shown to be decreased. The elevated serum PTHrP might, therefore, have a physiologically compensatory role on the calcium regulatory systems in NIDDM. Furthermore, this elevation is most likely due to the excess production of this peptide and not to the decrease in urinary excretion.</p>","PeriodicalId":6910,"journal":{"name":"Acta endocrinologica","volume":"129 6","pages":"519-24"},"PeriodicalIF":0.0,"publicationDate":"1993-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1530/acta.0.1290519","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"19101467","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"Regulation of bovine and ovine lymphocyte proliferation by progesterone: modulation by steroid receptor antagonists and physiological status.","authors":"V H Monterroso, P J Hansen","doi":"10.1530/acta.0.1290532","DOIUrl":"https://doi.org/10.1530/acta.0.1290532","url":null,"abstract":"<p><p>Lectin-stimulated sheep and cow lymphocytes were used to test whether inhibitors of classical steroid receptors block suppressive effects of progesterone and whether effects of progesterone vary with physiological status. Neither RU 38486 nor RU 43044 blocked the inhibitory effects of progesterone on lymphocyte proliferation. Rather, these antagonists were themselves inhibitory. Effects of progesterone and antagonists were additive: the percentage inhibition caused by progesterone was similar whether antagonists were present or not. The degree of lymphocyte proliferation and the inhibitory effects of progesterone were of the same magnitude for pregnant/lactating cows, pregnant/non-lactating cows, postpartum/lactating cows and cyclic/non-lactating cows. In conclusion, progesterone does not appear to inhibit lymphocyte proliferation through actions that involve classical steroid receptors. There was no evidence that lymphocyte proliferation in the cow is suppressed during pregnancy or that the inhibitory effects of progesterone increase during pregnancy.</p>","PeriodicalId":6910,"journal":{"name":"Acta endocrinologica","volume":"129 6","pages":"532-5"},"PeriodicalIF":0.0,"publicationDate":"1993-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1530/acta.0.1290532","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"19101469","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
K Koike, K Kadowaki, K Hirota, M Ohmichi, H Ikegami, T Sawada, A Miyake, O Tanizawa
{"title":"Prolactin stimulates [3H]dopamine release from dispersed rat tubero-infundibular dopaminergic neurons and dopamine decreases gonadotropin-releasing hormone release induced by calcium ionophore.","authors":"K Koike, K Kadowaki, K Hirota, M Ohmichi, H Ikegami, T Sawada, A Miyake, O Tanizawa","doi":"10.1530/acta.0.1290548","DOIUrl":"https://doi.org/10.1530/acta.0.1290548","url":null,"abstract":"<p><p>In order to investigate the involvement of prolactin-dopamine and dopamine-gonadotropin interactions in the hypothalamo-pituitary axis of hyperprolactinemia, in vitro studies were performed using primary cultures of dispersed rat hypothalamic heterogeneous cells containing tubero-infundibular dopaminergic neurons or gonadotropin-releasing hormone (GnRH) neurons. We observed that prolactin caused dose-dependent stimulation of [3H]dopamine release after a 16-h incubation. Staurosporin (10 nmol/l), an inhibitor of protein kinase C, significantly reduced the [3H]dopamine release induced by prolactin (1 mg/l). Incubation of tubero-infundibular dopaminergic neurons with prolactin (1 mg/l) had no effect on intracellular cyclic adenosine monophosphate accumulation. Dopamine (1 mumol/l) significantly (p < 0.01) reduced the release of GnRH induced by 50 mumol/l calcium ionophore from dispersed hypothalamic cells from the preoptic area, while prolactin had no effect on GnRH release. These data support the hypothesis that the antigonadotropic effect of prolactin on the hypothalamus is mediated by an inhibitory effect of dopamine on GnRH release.</p>","PeriodicalId":6910,"journal":{"name":"Acta endocrinologica","volume":"129 6","pages":"548-53"},"PeriodicalIF":0.0,"publicationDate":"1993-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1530/acta.0.1290548","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"19101472","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"Plasma glucagon responses to insulin-induced hypoglycemia and arginine in spontaneous non-insulin-dependent diabetes mellitus (NIDDM) rats, Otsuka Long Evans Tokushima Fatty (OLETF) strain.","authors":"K Ishida, A Mizuno, T Sano, K Shi, K Shima","doi":"10.1530/acta.0.1290585","DOIUrl":"https://doi.org/10.1530/acta.0.1290585","url":null,"abstract":"<p><p>Pancreatic A-cell function in the newly developed Otsuka Long Evans Tokushima Fatty (OLETF) strain of non-insulin-dependent diabetes mellitus (NIDDM) rats was examined in relation to the morphological changes in their islets and the plasma glucagon responses to insulin-induced hypoglycemia and an arginine test by chronological studies in seven male OLETF and seven male non-diabetic control Long Evans Tokushima Otsuka (LETO) rats each at 10, 16 and 24 weeks of age and eight male OLETF rats that were placed in a cage with a wheel for exercising from 5 to 24 weeks of age. The hormonal contents and morphological features of the pancreas of these rats were examined. After iv injection of insulin, the plasma glucagon level rose significantly from the basal level in OLETF rats at 10 weeks old, but little if at all in those of 16 and 24 weeks old. The pancreatic A cells of LETO rats of all age groups responded equally well to glucopenia. The areas under the response curves of plasma glucagon (sigma delta IRG) during the 90 min of insulin-induced hypoglycemia were 14496 +/- 7860 vs 9588 +/- 3930, 2257 +/- 3018 vs 9235 +/- 5447 (p < 0.05) and 826 +/- 985 vs 9707 +/- 2510 (p < 0.01) ng.min-1.l-1 in OLETF rats vs LETO rats of 10, 16 and 24 weeks old, respectively. The plasma glucagon responses during the arginine test were higher in OLETF rats than in LETO rats at 10 and 16 weeks but not at 24 weeks of age.(ABSTRACT TRUNCATED AT 250 WORDS)</p>","PeriodicalId":6910,"journal":{"name":"Acta endocrinologica","volume":"129 6","pages":"585-93"},"PeriodicalIF":0.0,"publicationDate":"1993-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1530/acta.0.1290585","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"19102067","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}