circ_0072088 promotes the malignant biological behaviors of thyroid cancer IHH4 cells by regulating the miR-532-3p/WEE1 axis / 中国肿瘤生物治疗杂志

@#[摘 要] 目的：探讨circ_0072088对甲状腺癌IHH4细胞恶性生物学行为的影响及其可能的机制。方法：选取2015年4月至2019年3月于青海大学附属医院就诊的确诊为甲状腺癌后接受切除手术且术前未进行任何治疗的48例甲状腺癌患者的甲状腺癌组织及其对应癌旁组织。根据GEO数据集（GSE93522）分析鉴定甲状腺乳头状癌中差异表达的circRNA。用qPCR法检测circ_0072088在甲状腺癌组织和细胞中的表达水平。采用CCK-8法和Transwell法检测circ_0072088和miR-532-3p过表达对甲状腺癌IHH4细胞增殖、迁移和侵袭的影响。采用生物信息学和双荧光素酶报告基因实验验证甲状腺癌细胞中circ_0072088与miR-532-3p、miR-532-3p和WEE1基因之间的关系。结果：circ_0072088在甲状腺癌组织和细胞中呈高表达（P<0.05或P<0.01）。circ_0072088过表达促进了甲状腺癌细胞的增殖、迁移和侵袭（P<0.05或P<0.01），而转染 miR-532-3p模拟物能够减弱这种作用（P<0.05或P<0.01）。此外，机制研究表明，circ_0072088可以与miR-532-3p靶向结合，且WEE1是miR-532-3p的下游靶基因。结论：circ_0072088通过调控miR-532-3p/WEE1轴促进IHH4细胞的增殖、迁移和侵袭。

Expression of SHH and IHH during the formation of intervertebral discs in mouse embryo / 中国组织工程研究

BACKGROUND: Studies have shown that Sonic Hedgehog (Shh) is expressed in mouse Notochordal cells, and India Hedgehog (Ihh) is mainly expressed in chondrocyte-like cells and end plates of mouse embryonic vertebral bodies. However, the expression of Shh and Ihh during the formation of intervertebral discs in mouse embryo is unclear. OBJECTIVE: To investigate the expression of Shh and Ihh during the formation of intervertebral discs in mouse embryo. METHODS: Male Shh-CreERT2, R26-mTmG/+ mice mated with female R26-mTmG/+ mice to obtain pregnant rats at different periods of pregnancy (E8.5, E11.5, E12.5, E14.5, E16.5, E18.5), followed by injection of 10 g/L tamoxifen at a dose of 10 μL/g. Intervertebral disc tissues were isolated from the mouse embryo at P0, followed by genotype identification. At the same time, male C57BL/6 mice mated with female C57BL/6 mice to obtain pregnant rats at different periods of pregnancy (E11.5, E12.5, E14.5, E16.5, E18.5), and the intervertebral disc tissues of mouse embryo were taken and analyzed. The expression of Shh and Ihh during the formation of intervertebral discs in mouse embryo was detected using immunofluorescence and immunohistochemistry, respectively. Implementation of animal experiments was approved by the Animal Ethics Committee of Soochow University. RESULTS AND CONCLUSION: Shh-CreERT2; R26-mTmG/+ mouse embryos were identified by PCR amplification. The results of immunofluorescent staining showed a gradual decrease in the expression of Shh in nucleus pulposus cells during the formation of intervertebral discs in mouse embryo. The results of immunohistochemical staining showed a gradual increase in expression of Ihh in nucleus pulposus cells during the formation of intervertebral discs in mouse embryo. In summary, Shh and Ihh are dynamically expressed during the formation of intervertebral discs in mouse embryo, and provide a basis for further research on the molecular mechanism of intervertebral disc degeneration.

Effects of fluoride on the expression of Indian hedgehog, parathyroid hormone-related peptide, smoothened protein and mRNA and on proliferation and apoptosis of chondrocytes / 中华地方病学杂志

Objective To study the proliferation and apoptosis and investigate the expression of Indian hedgehog (Ihh),parathyroid hormone-related peptide (PTHrp),smoothened (Smo) protein and mRNA in the cultured rat primary chondrocytes exposed to different doses of NaF.Methods The third generation articular chondrocytes of neonate rat were cultured in vitro and treated with 0 (control),5,10,20 and 40 mg/L of fluoride.The proliferation activities of cells at different times (24,48 and 72 h) were tested by Thiazolyl Blue Tetrazolium Bromide (MTT).The apoptosis rate was determined by flow cytometry.The expressions of protein and mRNA of Ihh,Smo and PTHrp at 48 h were determined by Western blotting and semi-quantitative RT-PCR,respectively.Results After exposed to 5 mg/L of fluoride for 24,48 and 72 h,the proliferation rates were significantly increased [(1.17 ± 0.07)％,(1.20 ±0.06)％,(1.16 ± 0.08)％] compared with those of control group [(1.10 ± 0.08)％,(1.13 ± 0.08)％,(1.15 ± 0.08)％],but the proliferation activity at 48 and 72 h in 40 mg/L group [(0.72 ± 0.11)％,(0.68 ± 0.04)％] was significantly lower than those in control group (all P ＜ 0.05).Compared with the control group,apoptosis rate of cartilage cell in fluoride treatment group increased gradually [(1.47 ± 0.05)％,(19.87 ± 3.03)％,(25.30 ± 1.28)％,(45.73 ± 4.63)％,F =123.328,P ＜ 0.01].Western blot analysis and RT-PCR results showed that the Ihh,PTHrp,Smo mRNA and protein expression increased in the fluoride groups at 48 h (Ihh protein:0.77 ± 0.08 vs.0.98 ±-0.07,1.23 ± 0.06,1.37 ±0.07,1.34 ± 0.07;PTHrp protein:0.68 ± 0.04 vs.0.89 ± 0.05,0.83 ± 0.05,1.29 ± 0.05,1.16 ± 0.08;Smo protein:0.37 ± 0.01 vs.0.64 ± 0.06,0.67 ± 0.03,0.96 ± 0.06,0.69 ± 0.06;Ihh mRNA:0.77 ± 0.05 vs.0.98 ± 0.05,1.09 ±0.05,1.27 ± 0.03,1.46 ± 0.06;PTHrp mRNA:0.67 ± 0.07 vs.0.97 ± 0.05,1.07 ± 0.08,1.37 ± 0.05,1.45 ± 0.05;Smo mRNA:0.45 ± 0.03 vs.0.63 ±-0.04,0.71 ± 0.05,0.81 ± 0.01,1.00 ± 0.02,all P ＜ 0.05).Conclusions Low doses of fluoride can promote the proliferation of chondrocytes cultured in vitro,and high doses of fluoride can promote the apoptosis of chondrocytes cultured in vitro.The expression of Ihh signaling pathway RNAs and proteins of the cartilage cells are increased following increased levels of fluoride.The results suggest that fluorine has activated the Ihh signaling pathway in chondrocytes and promoted the proliferation and apoptosis processes which might be involved in chondrocytes injury.

Displasia septo-optica. Evaluación clínica, endocrinológica y neuro-radiológica. Seguimiento de 46 pacientes pediátricos / Septo-optic dysplasia. Clinical, endocrinological, and neuroradiological evaluation. Follow-up of 46 pediatric patients

La displasia septo-óptica (DSO) es una condición rara y altamente heterogénea, definida por la combinación de hipoplasia del nervio óptico (HNO), malformaciones cerebrales de la línea media, tales como aplasia/hipoplasia de septum pellucidum y cuerpo calloso, e insuficiencia hipotálamo-hipofisaria de grado variable. Se realizó un trabajo que tuvo como objetivo caracterizar la población de pacientes con diagnóstico de DSO seguidos en nuestro Hospital durante 7 años. Se incluyeron 46 pacientes (18 mujeres) que fueron divididos en 2 grupos, según tuviesen o no insuficiencia hipotálamo-hipofisaria (IHH). El 58.7% (n=27) presentó IHH de algún tipo, mientras que el 41.3% (n=19) no la presentó. En aquellos 19 pacientes con IHH se diagnosticaron deficiencia de GH y TSH (85.1%) y de ACTH (48.1%). La longitud corporal (mediana) del grupo con IHH fue más baja (p = 0,01) que la del grupo sin IHH, a pesar de que la edad fue menor a 2 años en todos los casos. Los pacientes fueron seguidos 1,3-8,3 años. Se observaron incidencias similares de agenesia del cuerpo calloso, del septum pellucidum, y ventriculomegalia, pero las alteraciones del desarrollo cortical se observaron con mayor frecuencia en los pacientes sin IHH. La ictericia neonatal, convulsiones y/o hipoglucemia, y micropene en neonatos y lactantes con DSO se presentaron en el subgrupo con IHH. El 58,7% de los pacientes con DSO presentaron algún grado de insuficiencia hipotálamo-hipofisaria. En la mayoría de los casos el diagnóstico de IHH no se realizó en el momento de aparición de los síntomas, sino más tardíamente en su seguimiento. En el 45% de los pacientes se evaluaron alteraciones radiológicas del SNC, específicamente en la región hipofisaria. Una fracción importante de las deficiencias de TSH/T4 (36,4%), GH (50%) y ACTH (23%) aparecieron mas tardíamente en el curso de la evolución. En 10 niños con déficit de hormona de crecimiento (2 tests farmacológicos sin respuesta) se realizó el tratamiento sustitutivo con rhGH (durante un periodo de 4±3 años), observándose una mejoría promedio de + 1,5 SDS en la talla de estos pacientes. En conclusión, la hipoplasia neonatal de nervios ópticos, asociada o no a ictericia e hipoglucemia, debe ser un signo de alarma para el diagnóstico de DSO, con riesgo de insuficiencia suprarrenal, shock y muerte, y puede requerir, por lo tanto, urgente tratamiento. Las deficiencias pueden aparecer en el curso de la evolución, a pesar del carácter congénito de la anomalía. Finalmente, se deben sustituir las deficiencias hormonales y tener presente que el tratamiento con rhGH puede mejorar la talla final en estos pacientes (AU)

Septo-optic dysplasia (SOD) is a rare and highly heterogeneous condition consisting of a combination of optic nerve hypoplasia (ONH), midline brain abnormalities, such as aplasia/hypoplasia of the septum pellucidum (ASP) and corpus callosum; and variable degree of hypoyalamo-pituitary insufficiency. The aim of this study was to characterize a population of SOD patients diagnosed and followed at the Garrahan Pediatric Hospital, from 1989 to 2006. We included 46 patients (18 females), that were divided into two groups according to the presence or absence of hypothalamic-pituitary insufficiency (IHH). Fifty nine% of SOD patients presented with IHH. GH and TSH deficiencies were diagnosed in 85.1% of IHH patients, while ACTH deficiency was found in 48.1%. Height (median) for the IHH group was shorter (p = 0,01) than for the group without IHH. Patients were followed for 1.3-8.3 years. Similar incidence of corpus callosum and/or septum pellucidum agenesis and ventriculomegaly were found in the two groups, but we observed more association with cortical developmental disorders in patients without IHH. In newborns, the association of ophthalmologic disorders and jaundice, seizures and/or hypoglycemia and micropene should frequently lead to the diagnosis of SOD and IHH. While 58,7% of DSO patients presented with hypothalamic-pituitary deficiency, only 45% of them showed sellar radiological abnormalities. Although SOD is a congenital disease, hormonal deficiencies may appear during follow-up. In 10 children with SOD and GH deficiency, rhGh treatment (for 4±3 years) improved height in 1.5 SDSs. In conclusion: in newborns with nerve optic hypoplasia, associated or not with jaundice, seizures and hypoglycaemia, the diagnosis of SOD and IHH should be considered. Treatment could be an emergency need because of risk of adrenal insufficiency and hypoglycemia (AU)

Severe osteoporosis secondary to idiopathic hypogonadotrophic hypgonadism (IHH) in a 45-year old male

SYNOPSIS: Male hypogonadi sm i s an impor tant and treatable cause of osteoporosis. Severe osteoporosis leading to multiple osteoporotic fractures from idiopathic hypogonadotrophic hypogonadism (IHH) is rare. The present case illustrates the significance of timely and thorough evaluation of young adult males presenting with a seemingly ordinary complaint of "bone pains."THE CASE: I report a case of a 45 year-old male presenting with a 6-year history of progressive bone pains. Most prominent laboratory findings include low total serum tes tos terone (4.6 nmol/L) in the background of an inappropriately normal serum FSH, LH and sex hormonebinding globulin (SHBG). There is associated elevated urinary N- telopeptide (4x). Sperm analysis showed oligospermia. Scrotal ultrasound revealed normal-sized descended testis with no solid masses. Skeletal survey showed generalized decrease in bone density. Dual energy x-ray absortiometry (DXA) showed severe osteoporosis. Cranial CT scan with contrast did not show a sellar-suprasellar mass.TREATMENT AND OUTCOME: The patient was diagnosed with severe osteoporosis secondary to IHH. The patient received zoledronic acid (Aclasta) 5mg IV infusion. Two months after discharge, the patient reports a significant decrease in bone pains leading to more mobility. He is scheduled for his first dose of a GnRH agonist (Leuprodin acetate 3.75mg IM) to induce testosterone production.DISCUSSIONS: The incidence of osteoporosis among males is indirectly correlated to the reduction in circulating testosterone. First - line t reatment of os teoporosis in hypogonadal men is with bisphosphonates. Bisphosphanate therapy increase BMD, reduces vertebral fracture risk and is currently considered the standard of care for osteoporotic care for men.CONCLUSION: Osteoporosis is fast becoming a common condition among males. Osteoporotic fractures are associated with substantial morbidity and mortality. The present case emphasizes the importance of thorough and timely evaluation among men with low BMD or low-trauma fractures, which should include laboratory assessment to exclude secondary causes such as hypogonadism.