Progesterone and estrogen receptor expression by canine cutaneous soft tissue sarcomas / Expressão de receptores para progesterona e estrógeno em sarcomas de tecidos moles cutâneos em cães

Canine soft tissue sarcomas (STS) comprise a heterogeneous group of malignancies that share similar histopathological features, a low to moderate recurrence rate and low metastatic potential. In human medicine, the expression of estrogen receptors (ER) and progesterone receptors (PR) in sarcomas has been studied to search for prognostic factors and new treatment targets. Similar studies have yet to be conducted in veterinary medicine. The objective of this study was therefore to investigate by immunohistochemistry (IHC) the ER and PR expression in a series of 80 cutaneous and subcutaneous sarcomas in dogs with histopathological features of peripheral nerve sheath tumor (PNST) and perivascular wall tumor (PWT). All cases were positive for PR and negative for ER. Tumors of high malignancy grade (grade III) exhibited higher PR expression than low-grade tumors (grade I). Tumors with mitotic activity greater than 9 mitotic figures/10 high power fields also exhibited higher PR expression. In addition, there was a positive correlation between cell proliferation (Ki67) and PR expression. Therefore, it is possible that progesterone plays a greater role than estrogen in the pathogenesis of these tumors. Future studies should explore the potential for selective progesterone receptor modulators as therapeutic agents in canine STS, as well as evaluating PR expression as a predictor of prognosis.(AU)

Sarcomas de tecidos moles (STM) caninos compreendem um grupo heterogêneo de neoplasias malignas, que apresentam alterações histopatológicas similares, baixa a moderada taxa de recorrência e baixo potencial metastático. Em medicina humana, a expressão de receptor para estrógeno (RE) e receptor para progesterona (RP) nos sarcomas tem sido estudada, visando a busca por fatores prognósticos e novos alvos para tratamentos. Na medicina veterinária, ainda não foram realizados estudos similares. O objetivo deste trabalho foi investigar por imuno-histoquímica a expressão de RE e RP em uma série de 80 sarcomas cutâneos e subcutâneos de cães, com características histopatológicas de tumor de bainha de nervo periférico e tumor de parede perivascular. Todos os casos foram positivos para RP e negativos para RE. Tumores de alto grau de malignidade (grau III) exibiram maior expressão deste receptor que os tumores de baixo grau (grau I). Tumores com atividade mitótica maior que 9 figuras mitóticas/10 campos de grande aumento também exibiram maior expressão do RP. Em adição, houve correlação positiva entre o índice de proliferação celular (Ki67) e a expressão de RP. Assim, é possível que a progesterona desempenhe maior papel que o estrógeno na patogênese desses tumores. Futuros trabalhos poderão explorar o potencial dos moduladores seletivos de RP como agente terapêutico em STM caninos, bem como avaliar a expressão de RP como preditiva de prognóstico.(AU)

Effect of Acupuncture Combined with Gavage of Rule Granule on Serum Prolactin, Estradiol and Progestone Contents and Their Receptor Expression in Breast in Mammary Gland Hyperplasia Rats / 针刺研究

OBJECTIVE: To observe the effect of acupuncture combined with medication (Rule Granule) on serum prolactin (PRL)，estradiol (E2) and progestone (P) contents and expressions of prolactin receptor (PRLR), estrogen receptor (ER) and progestrone receptor (PR) proteins in mammary gland (MG) tissues of MG hyperplasia (MGH) rats, so as to explore its mechanisms underlying improvement of MGH. METHODS: A total of 55 female SD rats were randomized into blank control group (n＝10), model group (n＝10), acupuncture group (n＝11), medication group (n＝10), and acupuncture plus medication group (n＝9). The MGH model was established by muscular injection of E2 benzoate(0.5 mg/kg, once daily for 20 days), followed by injection of P (5 mg/kg) into the lateral muscle of the hind-limb, once daily for 5 days, and the rats of the control group were treated by muscular injection of normal saline at the same site and in the same procedures. Acupoint group A composed of bilateral Tianzhong (SI 11), Ganshu (BL 18) and Zusanli (ST 36), and group B composed of Tanzhong (CV 17), and bilateral Wuyi (ST 15) and Hegu (LI 4) were alternatively punctured with filiform needles and stimulated swiftly by twirling the needle in each acupoint for 20－30 sec, once daily for 30 days. Rats of the medication group and acupuncture ＋medication group were treated by gavage of Rule Granule fluid containing Baishao (Paeonia Iactiflora Pall), Danggui (Angelica sinensis), etc. (1.5 mL/100 g for each rat) and those of the other groups treated by gavage of distilled water (1.5 mL/100 g). The treatment was given once daily for continuous 30 d. At the end of the experiments, the rats' abdominal aorta blood was collected for assaying the contents of serum PRL, E2 and P with ELISA, and the pathological changes of breast tissue (the left 2nd pairs) were observed under microscope after sectioning and H．E. staining. The expression of PRLR, ER and PR proteins in the breast tissue was detected by immunohistochemistry. RESULTS: After modeling, the hyperplasia of mammary gland in the model group was obvious; the serum PRL and E2 contents and the immunoactivities of PRLR, ER and PR were significantly increased (P0.05)．. CONCLUSION: Acupuncture, Rule Granule, and acupuncture combined with Rule Granule can improve hyperplasia of mammary gland in rats, which may be related to their effects in reducing serum PRL and E2 and breast ER, PR and PRLR expression levels, and in increasing serum P level. The therapeutic effects of acupuncture plus Rule Granule are better than those of simple acupuncture and simple Rule Granule.

Hormones and gallbladder cancer in women.

Epidemiological evidence suggests that the incidence of gallstone disease and gallbladder cancer is higher in women. We analyzed the literature on estrogen and progesterone receptor expression in gallbladder cancer in women. A systematic search was done using Medline, Embase, and Cochrane Central Register of Controlled Trials for the years 1983–2009. The search terms used included ‘gallbladder’, ‘gallstone’, ‘oestrogen/estrogen’, ‘progesterone’, ‘cancer’, ‘cholelithiasis’, ‘hormone,’ and ‘motility’. Hormone receptor expression in gallbladder cancer was analyzed in 11 studies of which immunohistochemistry was used in 10 and enzyme immunoassay in one study. Sample sizes varied from 3 to 141. Estrogen and/or progesterone receptor expression was detectable in gallbladder cancer tissue samples in nine studies, whereas four studies failed to confirm these findings. The data on the association of hormone receptor expression to tumor differentiation is contradictory and needs further evaluation.

Differential Expression of Ryanodine Receptors in the Developing Rat Cochlea / 听力学及言语疾病杂志

Objective Ryanodine receptor (RyR) is one of the Ca2+ release channels on the intracellular Ca2+ stores. RyR induced-Ca2+ release is activated by the voltage-dependent Ca2+ entry, that is, calcium-induced calcium release (CICR). Intracellular free Ca2+ concentration (ECa2+]i) plays a key role on cochlear function. Our study is to investigate the differential expression of RyR in the developing rat cochlea, and to analyze the relationship between the expression of RyR and auditory functional development. Methods Immature SD rats, which were 1 day (P1), 5 days (P5), 10 days (P10), 14 days (P14), 28 days (P28) after parturition, and adutl rats(5 rats for each age) were included in the study. Frozen cochlea sectioning and immunofluorescence were applied to observe the differential expression of RyR. Results The RyR expression in the Corti's organ increased during the cochlear development. It's not significant that the stain was observed on the hair cells and supporting cells in the Corti's organ of P1 and P5 rats. The appearance of the Corti's organ of P10 rats trended to maturity. In P14 rats the RyR expression on hair cells located in the synaptic area against the afferent or efferent nerve, and the strain on supporting cells was extensive. There was little different between the strain on cochlea of P14 and P28 rats. In postmature rat spiral ganglion neurons (SGN), the RyR expression verged gradually from extensive whole cell soma to the synaptic area near to the plasma membrane. Conclusion The RyR expression peaked the 14th day after parturition, which was close to that in the mature cochlea. The time course of the RyR expression during the cochlear development was coincident with that of the auditory functional development. The RyR expression on both hair cells and SGNs located in the synaptic area near to the plasmolemma, implying that RyR induced-CICR was related to the auditory functions such as neurotransmission. Extensive RyR expression in the soma of SGNs at the early stage possibly involved in apoptosis of SGNs during neuron development.

Recurrencia y supervivencia en cáncer mamario temprano con inmunofenotipo triple-negativo / Recurrence and survival rates among early breast cancer cases with triple negative immunophenotype

Antecedentes: El estado ganglionar axilar, la expresión de los receptores hormonales y del HER2 son importantes factores pronóstico en cáncer de mama temprano. El inmunofenotipo triple negativo (HER2 y receptores hormonales negativos) se ha asociado con mayor frecuencia de recurrencia y menor tiempo de supervivencia. El objetivo de esta investigación fue evaluar el comportamiento clínico, recurrencia y supervivencia en mujeres con cáncer de mama temprano-triple negativo y otros inmunofenotipos. Material y métodos: Estudio retrospectivo de mujeres en etapas IIIB, mayores de 18 años, en quienes se determinó la expresión de la proteína HER2, receptores de estrógeno y de progesterona a través de inmunohistoquímica. Se identificaron cinco grupos: triple negativo, triple positivo, HER2 negativo y receptores hormonales positivos, HER2 positivo y receptores hormonales negativos, HER2 negativo y un receptor hormonal positivo. En cada caso se analizó la edad, fecha del diagnóstico, etapa clínica, tamaño tumoral, estado ganglionar axilar, receptores de estrógenos, progesterona, HER2, p53, angiogénesis, Ki67, tipo de cirugía realizada, tratamiento adyuvante, tiempo a la recurrencia, número y sitios de la recurrencia, así como el tiempo de sobrevida global. Resultados: 17 pacientes (15.4%) manifestaron el fenotipo triple negativo; 14 (12.7%), triple positivo; 52 (47.3%) en el grupo 3, 11 (10%) en el 4 y 16 (14.5%) en el grupo 5. El fenotipo triple negativo se asoció con proliferación celular aumentada (p<0.000), menor edad (mediana 43 años), mayor tamaño tumoral (mediana 2.5 cm) y menor proporción de pacientes en etapa I, así como mayor frecuencia de expresión positiva de la proteína p53 (78.5%). Observamos mayor frecuencia de recurrencia y de muerte en el grupo triple negativo y en HER2 positivo con receptores hormonales negativos. Conclusiones: El cáncer de mama triple negativo se presenta en mujeres jóvenes y se asocia con proliferación celular aumentada, induce mayor incidencia de recurrencia y de mortalidad. El comportamiento biológico del cáncer de mama con fenotipo triple negativo es agresivo y similar al observado en pacientes con HER2 positivo y receptores hormonales negativos.

BACKGROUND: Axillary lymph node status, hormonal receptors (HR) and HER2 expression are significant prognostic factors for early breast cancer. Triple negative immunophenotype (HER2 and HR negative) is associated with a high frequency of recurrence and lower overall survival. The objective was assess clinical behavior, recurrence and survival of patients with triple negative early breast cancer and patients with other immunophenotypes. MATERIAL AND METHODS: We carried out a retrospective study among women with stages I-IIB over 18 years with determination of HR and HER2 expression by immunohistochemical assay. We identified 5 groups: triple negative, triple positive, HER2 negative & HR positive, HER2 positive & HR negative, HER2 negative & 1 HR positive. We recorded age, date of diagnosis, clinical stage, tumor size, axillary lymph node status, ER, PR, HER2, p53, angiogenesis, Ki67, type of surgery, adjuvant treatment, time to recurrence, number and recurrence site and overall survival. RESULTS: 17 patients (15.4%) had triple negative phenotype, 14 (12.7%) triple positive, 52 (47.3%) were localized in group 3, 11 (10%) in 4 and 16 (14.5%) in group 5. Triple negative phenotype was associated with increased cellular proliferation (p < 0.000); being young (median 43 years), large tumor size (median size 2.5 cm) lower proportion of patients in stage I and high frequency of p53 positive (78.5%). We observed a high frequency of recurrence and death among the triple negative group and among the HER2 positive and HR negative cases. CONCLUSIONS: Triple negative breast cancer is more common among young women and is associated with a high frequency of recurrence and mortality. Clinical behavior among triple negative breast cancer cases is aggressive and displays a similar clinical profile that observed among HER2 positive and HR negative patients.

Requirements for in vitro growth of human thymocytes

Since it is difficult to study human thymocyte maturation in vitro, we have developed an in vitro thymocyte culture system which has allowed us to select the optimal growth conditions for thymocyte subpopulations. Three thymocyte subpopulations (CD3-CD1-, CD1+CD3-, and CD3+CD1-) were isolated by a single step percoll density gradient centrifugation and indirect panning procedure using anti-CD1 and anti-CD3 monoclonal antibodies, and their purity was checked by flow cytometry. The combination of concanavalin A (Con A), tetradecanoylphorbol acetate (TPA), and IL-2 was shown to be the most reliable stimulus for the proliferation of CD3-CD1- thymocytes for up to 15 days in a culture system in vitro. Flow cytometric analysis for the phenotypic change of CD3-CD1- thymocytes revealed a steady increase of CD3 antigen after a 3-day cultivation, whereas there was no change in CD1 antigen intensity. A combination of Con A and IL-2 was both sufficient and necessary to induce growth of CD3+CD1- thymocytes. The major population of immature cortical thymocytes (CD3-CD1+ or CD3+CD1+), which are considered to be the most unresponsive dead-end cells, could not be maintained or stimulated with any combination used in this experiment, even in the presence of thymic accessory cells.