RESUMEN

A novel heterometallic metal-porphyrinic framework (MPFs) built from Y and K ions as nods and meso-tetra(4-carboxyphenyl)porphyrin as linkers has been successfully synthesized and characterized. The single crystal X-ray diffraction indicated that this complex 1 exhibited a bilayered architecture of the porphyrins, which is seldom seen in MPFs. In addition, in vitro anticancer activity of complex 1 on three human breast cancer cells (BT474, SKBr-3 and ZR-75-30) was further determined.

Asunto(s)

Humanos , Porfirinas/química , Neoplasias de la Mama/tratamiento farmacológico , Estructuras Metalorgánicas/farmacología , Estructuras Metalorgánicas/química , Antineoplásicos/farmacología , Antineoplásicos/química , Valores de Referencia , Sales de Tetrazolio , Reproducibilidad de los Resultados , Cristalografía por Rayos X , Línea Celular Tumoral , Complejos de Coordinación/farmacología , Complejos de Coordinación/química , Formazáns

RESUMEN

A novel heterometallic metal-porphyrinic framework (MPFs) built from Y and K ions as nods and meso-tetra(4-carboxyphenyl)porphyrin as linkers has been successfully synthesized and characterized. The single crystal X-ray diffraction indicated that this complex 1 exhibited a bilayered architecture of the porphyrins, which is seldom seen in MPFs. In addition, in vitro anticancer activity of complex 1 on three human breast cancer cells (BT474, SKBr-3 and ZR-75-30) was further determined.

Asunto(s)

Antineoplásicos/química , Antineoplásicos/farmacología , Neoplasias de la Mama/tratamiento farmacológico , Estructuras Metalorgánicas/química , Estructuras Metalorgánicas/farmacología , Porfirinas/química , Porfirinas/farmacología , Línea Celular Tumoral , Complejos de Coordinación/química , Complejos de Coordinación/farmacología , Cristalografía por Rayos X , Formazáns , Humanos , Enlace de Hidrógeno , Estructura Molecular , Valores de Referencia , Reproducibilidad de los Resultados , Sales de Tetrazolio

RESUMEN

Immunological mechanisms have been proposed to underlie the pathogenesis of recurrent spontaneous abortion (RSA). Vitamin D has a potent immunomodulatory effect, which may affect pregnancy outcome. The objective of this study was to investigate 25-hydroxyvitamin D [25(OH) D] concentration and vitamin D receptor (VDR) expression in the decidual tissues of RSA patients. Thirty women with RSA (RSA group) and thirty women undergoing elective abortion (control group) were recruited during 2016 from gynecology outpatient clinics. We measured 25(OH) D, interleukin (IL)-17, IL-23, transforming growth factor ß (TGF-ß), VDR and 1-α-hydroxylase (CYP27B1) in decidual tissues collected during the abortion procedure. In the RSA group, 25(OH) D and TGF-ß were significantly decreased while IL-17 and IL-23 were significantly increased compared with the control group. VDR expression was significantly decreased in the RSA group compared with the control group. Logistic regression analysis showed a significant negative correlation between 25(OH) D in decidual tissues and RSA. These results indicated that vitamin D concentrations in the decidua are associated with inflammatory cytokine production, suggesting that vitamin D and VDR may play a role in the etiology of RSA.

Asunto(s)

25-Hidroxivitamina D3 1-alfa-Hidroxilasa/análisis , Aborto Habitual/metabolismo , Decidua/química , Receptores de Calcitriol/análisis , Vitamina D/análogos & derivados , 25-Hidroxivitamina D3 1-alfa-Hidroxilasa/metabolismo , Aborto Habitual/etiología , Adulto , Femenino , Humanos , Interleucina-17/análisis , Interleucina-23/análisis , Modelos Logísticos , Embarazo , Tercer Trimestre del Embarazo , Receptores de Calcitriol/metabolismo , Factores de Riesgo , Estadísticas no Paramétricas , Factor de Crecimiento Transformador beta/análisis , Vitamina D/análisis , Vitamina D/metabolismo , Deficiencia de Vitamina D/complicaciones , Adulto Joven

RESUMEN

Borax, a boron compound and a salt of boric acid, is known to inhibit the growth of tumor cells. HepG2 cells have been shown to be clearly susceptible to the anti-proliferative effects of borax. However, the specific mechanisms regulating this effect are poorly understood. This study aimed to investigate the pathways underlying the growth inhibition induced by borax in HepG2 cells. The effects of borax on HepG2 cell viability were characterized using MTT. Apoptosis was also verified by annexin V/propidium iodide staining. JC-1 dye and western blotting techniques were used to measure mitochondrial membrane potential and p53, Bax, and Bcl-2 protein expression, respectively. Relevant mRNA levels were measured by qRT-PCR. Borax inhibited the proliferation of HepG2 cells in a time- and dose-dependent manner in vitro. The apoptotic process triggered by borax involved the upregulation of p53 and Bax and the downregulation of Bcl-2, which was confirmed by a change in the mitochondrial membrane potential. These results elucidate a borax-induced apoptotic pathway in HepG2 cells that involves the upregulation of p53 and Bax and the downregulation of Bcl-2.

Asunto(s)

Apoptosis/efectos de los fármacos , Boratos/farmacología , Citostáticos/farmacología , Proteínas Proto-Oncogénicas c-bcl-2/metabolismo , Proteína p53 Supresora de Tumor/metabolismo , Proteína X Asociada a bcl-2/metabolismo , Boratos/toxicidad , Citostáticos/toxicidad , Regulación hacia Abajo , Células Hep G2 , Humanos , Proteínas Proto-Oncogénicas c-bcl-2/genética , Proteína p53 Supresora de Tumor/genética , Proteína X Asociada a bcl-2/genética

RESUMEN

Although blue-white screening based on α-complementation has been widely used in the screening of genetically modified bacteria, only a single blue-white screening is typically not enough to eliminate false positives. Sometimes, a secondary blue-white screening for the target colonies is required. In this study, two methods were used to investigate the feasibility of secondary blue-white screening for target colonies on lysogeny broth (LB)-ampicillin agar plates. The first method consisted of covering the target colonies grown on LB-ampicillin plate medium with a sterilized filter paper soaked in a solution of 60 µL 20 mg/mL X-gal and 8 µL 20% IPTG. The second method was that blue and white colonies were randomly selected from the blue-white screening plate medium and then re-streaked onto the blue-white screening medium. The colonies were then treated by two methods and incubated at 37°C for 12 h. The results showed that some of the white colonies treated by the two methods showed results similar to the colonies grown on the blue-white screening medium. These results indicate that the target colonies grown on blue-white screening medium can still be used to carry out a secondary blue-white screening. Thus, a blue-white screening liquid was successfully developed. Using the blue-white screening liquid, false positives can be eliminated directly based on the color of the target colonies. This will greatly improve the screening efficiency of positive clones and has important practical implications.

Asunto(s)

Bacterias/genética , Recuento de Colonia Microbiana/métodos , Ingeniería Genética/métodos , Agar/química , Bacterias/crecimiento & desarrollo , Medios de Cultivo/química , Galactósidos/farmacología , Indoles/farmacología , Isopropil Tiogalactósido/farmacología

RESUMEN

Habitual abortion is associated with the altered expression of multiple genes. This study was carried out to investigate the relationship between expression of Toll-like receptor 4 (TLR4) and monocyte chemotactic protein 2 (MCP2 or CCL8) and habitual abortion. This was done by detecting and comparing their relative expression in peripheral blood and placental villi of patients and healthy fertile women. Based on our previous research, 85 subjects with habitual abortion (study group) and 40 healthy fertile women (control group), who were admitted to our hospital between June 2013 and December 2014, were enrolled in this study. After these subjects signed written informed consent, peripheral blood samples and villous tissues were collected, from which the total RNA was extracted. The expression of TLR4 and MCP2 was detected with quantitative reverse transcription-polymerase chain reaction, using GAPDH as a reference control. The expression of TLR4 and MCP2 in the peripheral blood and villous tissues of the study group was significantly higher than that of the control group (P < 0.05). A positive correlation was also observed between the changes in expression levels of TLR4 and MCP2. In conclusion, TLR4 and MCP2 expression correlated with the occurrence of habitual abortion. Detecting expression changes in TLR4 and MCP2 in the peripheral blood is a feasible method for predicting the occurrence of abortion in women of child-bearing age.

Asunto(s)

Aborto Habitual/genética , Quimiocina CCL8/biosíntesis , Receptor Toll-Like 4/biosíntesis , Aborto Habitual/patología , Adulto , Quimiocina CCL8/genética , Vellosidades Coriónicas/metabolismo , Femenino , Regulación de la Expresión Génica , Humanos , Embarazo , Receptor Toll-Like 4/genética

RESUMEN

In order to evaluate the effect of high-concentrate diet supplementation on milk protein content, six Holstein dairy cows were assigned into high-concentrate diet (HC) or low-concentrate diet (LC) groups (N = 3/group) for 50 days. With regard to milk protein, HC feeding significantly reduced the percentage of milk protein (P < 0.01), and milk protein yield also reduced. The milk somatic cell count numbers and N-acetyl-D-glucosaminidase activity was significantly higher (P < 0.01) in the HC group than in the LC group. A pre-column derivatization procedure of o-phthalaldehyde was used to analyze the milk amino acid profile, the contents of Asp, Gln, Ala, Ile, Leu, and Lys were significantly lower in milk (P < 0.05), but Arg and Phe were significantly higher (P < 0.05) in the HC group than in the LC group. The mRNA abundance for amino acid transporters SLC7A8, SLC7A10 (P < 0.05), SLC1A3 (P < 0.05), and SLC16A10 (P < 0.05) were decreased in the HC group. These data indicate that expression of amino acid transporters alters regulation of amino acid utilization and decreases milk quality in dairy cows.

Asunto(s)

Sistemas de Transporte de Aminoácidos/biosíntesis , Dieta , Proteínas de la Leche/biosíntesis , Leche , Sistemas de Transporte de Aminoácidos/genética , Alimentación Animal , Animales , Bovinos , Femenino , Expresión Génica , Lactancia/genética , Proteínas de la Leche/genética

RESUMEN

The role of microRNAs (miRNAs) in the regulation of mammalian reproduction has been demonstrated previously. However, only a few studies have assessed the role of miRNAs in the reproduction processes of sheep. The elucidation of miRNA expression profiles in the ovaries of different sheep breeds representing fecundity extremes will be useful in understanding the roles of miRNAs in sheep reproduction. In this study, two small RNA libraries were constructed from ovary tissue taken from Qira black sheep and Hetian sheep during the estrous period and then sequenced using the Solexa sequencing method. We obtained 9,565,212 and 9,563,426 high-quality reads from Qira black sheep and Hetian sheep, respectively. In total, 531 miRNAs, including 98 putative miRNAs, were identified. Among the conserved miRNAs, 125 known miRNAs were significantly differentially expressed in the Qira black sheep and Hetian sheep libraries, with 24 upregulated and 101 downregulated in the Hetian sheep compared to the Qira black sheep. Four differentially expressed miRNAs were analyzed using real-time quantitative PCR to validate the reliability of the Solexa sequencing results. These results provide a foundation for future research on the regulation of miRNAs in sheep fertility and enrich the sheep miRNA databases.

Asunto(s)

MicroARNs/biosíntesis , MicroARNs/genética , Ovario/metabolismo , Ovinos/genética , Animales , Femenino , Secuenciación de Nucleótidos de Alto Rendimiento/veterinaria , MicroARNs/análisis , Reacción en Cadena en Tiempo Real de la Polimerasa/veterinaria , Análisis de Secuencia de ARN/veterinaria , Transcriptoma

RESUMEN

Angiotensin-converting enzyme 2 (ACE2), a newly discovered carboxypeptidase in the renin-angiotensin system (RAS), antagonizes ACE activity and plays an active role during tissue injury. Yet the mechanism of its action is not well known. Using a streptozotocin (STZ)-induced renal injury rat model, we investigated the relationship between renal injury and the antagonism between ACE and ACE2. We assayed the levels of urea nitrogen, urine glucose, creatinine, and protein, Ace2, Ace, angiotensin II type 1 receptor (At1) and Mas receptor mRNA, and renal and plasma angiotensin II (Ang II) in STZ-treated and untreated rats. We also used histology and immunohistochemistry to assess glomerular injury and ACE2 glomerular and cortical expression. The amounts of urea nitrogen, urine glucose, creatinine, and protein were significantly higher in STZ-treated rats than in control rats (P < 0.01). There were significant pathological changes in the kidney upon STZ-treatment. Ace2 and Ace mRNA levels were significantly higher in STZ-treated rats than in control rats (P < 0.05 and P = 0.05, respectively). There was no significant difference in the Mas receptor and At1 mRNA levels in the 2 groups, although At1 levels showed an increase upon STZ-treatment. The Ang II level in the renal cortical tissue and plasma of STZ-treated rats was higher than that of control rats (P < 0.05). The increase in Ace mRNA levels was higher than that of Ace2 mRNA levels, leading to an elevated Ace/Ace2 ratio. Together, these data suggest that the ACE-Ang II-AT1 axis is the dominant axis in severe kidney injury.

Asunto(s)

Lesión Renal Aguda/enzimología , Peptidil-Dipeptidasa A/metabolismo , Sistema Renina-Angiotensina/genética , Lesión Renal Aguda/inducido químicamente , Lesión Renal Aguda/patología , Angiotensina II/metabolismo , Enzima Convertidora de Angiotensina 2 , Inhibidores de la Enzima Convertidora de Angiotensina/metabolismo , Animales , Peptidil-Dipeptidasa A/genética , Ratas , Estreptozocina/toxicidad

RESUMEN

Dioscorea opposita Thunb. has been used as health food and herbal medicinal ingredients in traditional Chinese medicine. In this study, the total DNA of D. opposita Thunb. was extracted using an improved cetyltrimethylammonium bromide (CTAB) method, and the extracted DNA was further used for random amplified polymorphic DNA (RAPD) reaction system by design of the L16 (4(4)) orthogonal diagram. The results showed that the improved CTAB method can be used to isolate high-quality and high-concentration DNA, and the optimized protocol can overcome the instability of RAPD reaction system. The knowledge stated here can be used to study the genetic diversity of D. opposita Thunb.

Asunto(s)

ADN de Plantas/aislamiento & purificación , Dioscorea/genética , Técnica del ADN Polimorfo Amplificado Aleatorio/métodos , Cotiledón/genética , Dioscorea/clasificación , Humanos

RESUMEN

Aloe, an important folk herbal drug, includes abundant polysaccharides and secondary metabolites, which make it difficult to isolate high-quality DNA or RNA. In this paper, one and two improved methods were used to isolate the genomic DNA and RNA from the leaf of aloe, respectively. The obtained samples presented good quality and integrity; thus, they could be further used for many downstream molecular experiments. These reported protocols for DNA and RNA extraction offered a valuable reference for other related studies.

Asunto(s)

Aloe/química , Fraccionamiento Químico/métodos , ADN de Plantas/aislamiento & purificación , ARN de Planta/aislamiento & purificación

RESUMEN

Dioscorea opposita Thunb., included in the genus Dioscorea of the family Dioscoreaceae, is an important herb with great edible and medicinal value. In this study, the total RNA from leaves of Lichuan Dioscorea opposita Thunb. was isolated by an improved Trizol method. The results showed that the RNA extracted by the improved Trizol method had good integrity, and the RNA could be used for down-stream molecular biology operations including reverse transcription-polymerase chain reaction.

Asunto(s)

Fraccionamiento Químico/métodos , Dioscorea/química , ARN de Planta/aislamiento & purificación , ARN/aislamiento & purificación

RESUMEN

DC and AC electrical conductivity of bionanocomposites based on the immiscible polymer blend poly(epsilon-caprolactone)/polylactide (PCL/PLA, w/w 70/30), loaded with multiwall carbon nanotubes (CNT), were studied in a wide frequency range, 10(-3) < or = f < or = 10(7) Hz from 143 to 313 K. The nanofiller concentration ranged from 0 to 4 wt % and it was shown to be selectively located in the PCL phase. The PCL crystallinity degree was not affected by the presence of CNT. The variation of the DC conductivity allowed the determination of the percolation threshold, p(c) = 0.98 wt %, and the critical exponent t = 2.2 of the scaling law. The linear dependence of log (sigma(DC)) versus p(-1/3) showed the existence of tunneling conduction among CNT not yet in physical contact. The temperature independent results indicated a conventional tunnel effect. The AC conductivity of the nanocomposites followed the predictions of the universal dynamic response and the s exponents were determined at low concentrations. Master curves are presented showing the length and temperature-time superpositions.

Asunto(s)

Nanotubos de Carbono , Poliésteres/química , Microscopía Electrónica de Transmisión