Abstract Introduction/objectives Systemic lupus erythematosus (SLE) is a classic prototype of the multisystem autoimmune disease and follows a relapsing and remitting course. Triptolide is a diterpene triepoxide extracted from Chinese medicine Tripterygium wilfordii Hook F, with potent immunosuppressive and anti-inflammatory properties. Our previous work observed that triptolide alleviated lupus in MRL/lpr lupus mice with the upregulation of regulatory T cells (Treg) proportion in previous study. In this study, we explored the proportion of follicular T regulatory (Tfr), follicular T helper (Tfh) and germinal center (GC) B cells in lupus mice and evaluated the efficacy of triptolide for lupus treatment in vivo. Methods 20 female MRL/lpr mice were randomly divided into 2 treatment groups and treated orally with vehicle or triptolide. C3H mice were all housed as controlled group and treated orally with vehicle. The percentage of Tfr cells, Tfh cells and GC B cells in spleen of mice were detected by Flow cytometric analysis and immunohistochemistry after 13 weeks of treatment. Results We found that the percentage of Tfr cells decreased in MRL/lpr mice compared with controlled mice. The percentage of Tfh cells in MRL/lpr mice was significantly higher compared with that in controlled mice. The ratio of Tfr/Tfh is also decreased in lupus mice. After treated with triptolide in MRL/Lpr mice in vivo, the percentage of Tfr cells and ratio of Tfr/Tfh increased. The proportion of GC B cells also decreased in mice treated with triptolide by FACS and immunohistochemistry. Conclusions Our results demonstrate that the effect of triptolide in alleviating lupus is partly by reversing immune imbalance with increased percentage of Tfr cells and ratio of Tfr/Tfh. Triptolide might also has effect on immune response through inhibiting proliferating GC B cells.

Abstract GnRH analogues were widely used for controlld ovary stimulation, but their effects on oocyte quality remain contradictory. This study aimed to explore the influence of GnRH analogues on oocyte quality in mice. A total of 120 mice were randomly assigned to four groups:(i)GnRH-a+PMSG group; (ii) GnRH-ant+PMSG group; (iii) PMSG group; (iv) Control group. Ovaries were collected for quantitative real-time polymerase chain reaction (qRT-PCR) to assess GDF9 and BMP15 mRNA expression, and protein expression were evaluated by western blotting. Moreover, embryo developmental progress in vitro and implantation rate in vivo were recorded. Compared with control group, both GDF9 mRNA and protein expressions were strengthened in PMSG group, but reduced in the presence of GnRH-a or GnRH-ant. The GnRH-a group exhibited decreased BMP15 mRNA expression compared to PMSG group, while the GnRH-ant group did not show the same pattern. BMP15 protein expression were not statisticlly different among the four groups. Notably, there was no statistically difference in the expression of these two factors between GnRH-a and GnRH-ant groups. The percentage of zygotes progressing to the 2-cell stage and percentage of 2-cell advancing to the blastocyst stage were similar in the PMSG group and control group. However, both the GnRH-a and GnRH-ant groups showed decreased embryos development rates compared to other two groups. The embryonic implantation rate in control group (53.3%) was higher than that in the GnRH-a and GnRH-ant groups (33.3% and 30.8%, P<0.05). The difference between the PMSG (45.0%) and GnRHa group was statistically significant (P value of 0.023), but not between the PMSG and GnRH-ant group (P value of 0.486). No statistical difference was confirmed between GnRH-a and GnRH-ant groups. Our findings shed light on the safety of GnRH analogues in ovary stimulation, and highlight the need for further research to establish optimal and effective controlled ovary stimulation protocol.

The terahertz (THz) absorption spectra of 4- and 5-hydroxysalicylic acid were measured by terahertz time-domain spectroscopy (THz-TDS) in the range of 0.4-2.8 THz. The unit cells of both were calculated by density functional theory (DFT), and the theoretical simulation and experimental data were basically in agreement. To further interpret the origin of the characteristic absorption peaks, vibration characteristics and interactions of the unit cell model were analyzed by using potential energy distribution (PED) and interaction region indicator (IRI) method. The analysis indicated that the absorption peaks of 4- and 5-hydroxysalicylic acid were mainly derived from the vibration mode of dihedral angle torsion, and were closely related to the hydrogen bonds. And the intermolecular weak interaction of both were mainly dominated in quantity by van der Waals interaction. The results show that this is an effective solution to identify isomers and explain the formation mechanism of characteristic absorption peaks by using THz-TDS and combining PED and IRI methods.

Abstract Immature Litsea mollis fruits (ILMFs) are widely used as cold appetizers and seasonings in China. The free and bound volatile components of ILMFs from five distinct regions of China were determined by gas chromatography-mass spectrometry (GC-MS) combined with headspace solid phase microextraction (SPME) in the present work. The results reveal sixty six free-form volatiles and fifty two bound-form volatiles in ILMFs and terpene hydrocarbons and oxygenated terpenes are the dominate components. In addition, sixty-seven volatile compounds are found in Litsea mollis fruit for the first time. The free α-pinene, eucalyptol, linalool, α,β-citral, geraniol and citronellal, and bound β-myrcene, α-citral, geraniol, citronellal and eugenol are the major odor contributors of ILMFs. Multivariate statistical analysis indicates that the free and bound volatiles profiles of ILMFs of Fujian and Guangxi in southern China are similar and those of Yunnan and Sichuan in southwestern China are similar. The odor profiles of all of the ILMFs are floral, fruity and herbaceous.

Abstract Salted duck egg white was desalted and hydrolyzed to produce angiotensin I-converting enzyme (ACE) inhibitory peptides. Single factor test and response surface design were performed to determine the best hydrolysis conditions: enzyme dosage 11875.99 U/g, substrate concentration 33.04 g/L and hydrolysis time 4 h. The fraction V (MW < 1 kDa), which exhibited the strongest ACE inhibitory activity, was characterized by HPLC-ESI-MS/MS. Eighty-three peptides were identified, and among them Ile-Leu-Lys-Pro, Ile-Asn-Ser-Trp, Ile-Arg and His-Pro-Ala were synthesized for further research. Ile-Leu-Lys-Pro exhibited the highest ACE inhibitory activity (IC50: 0.355 mM). The molecular docking studies revealed that nine amino acids contributed to stabilize the docking complex. The ACE inhibition of Ile-Leu-Lys-Pro and Ile-Asn-Ser-Trp were mainly attributed to Ile in N-terminal. The residues Glu362 and Ala332 were the important binding sites in molecular docking. This research expands the understanding of ACE inhibitory peptides from duck egg white as well as highlights an opportunity for recycling an otherwise discarded byproduct.

ABSTRACT Growing evidence suggests that parasite-infected prey is more vulnerable to predation. However, the mechanism underlying this phenomenon is obscure. In small mammals, analgesia induced by environmental stressors is a fundamental component of the defensive repertoire, promoting defensive responses. Thus, the reduced analgesia may impair the defensive ability of prey and increase their predation risk. This study aimed to determine whether coccidia infection increases the vulnerability to predation in root voles, Microtus oeconomus (Pallas, 1776), by decreased analgesia. Herein, a predator stimulus and parasitic infection were simulated in the laboratory via a two-level factorial experiment, then, the vole nociceptive responses to an aversive thermal stimulus were evaluated. Further, a field experiment was performed to determine the overwinter survival of voles with different nociceptive responses via repeated live trapping. The coccidia-infected voles demonstrated reduced predator-induced analgesia following exposure to predator odor. Meanwhile, pain-sensitive voles had lower overwinter survival than pain-inhibited voles in enclosed populations throughout the duration of the experiment. Our findings suggest that coccidia infection attenuates predator-induced analgesia, resulting in an increased vulnerability to predation.

RESUMO Introdução: A acupuntura tem sido amplamente utilizada para alívio de dor. No entanto, seus mecanismos são muito pouco conhecidos. Objetivo: Investigar a relação entre o efeito analgésico da eletroacupuntura e a regulação da expressão de adenosina e de substância P. Métodos: Utilizou-se um modelo de dor inflamatória crônica em ratos por injeção única do Adjuvante Completo de Freund e, em seguida, os animais foram tratados com eletroacupuntura diariamente. Foram aplicadas sete sessões bilaterais de eletroacupuntura (ST36 e BL60, 0,5 a 1,5 mA, força inicial de 0,5 mA, aumentada em 0,5 mA a cada 10 minutos, 30 minutos por sessão, uma sessão por dia) em ratos com Adjuvante Completo de Freund, por sete dias. O efeito analgésico da eletroacupuntura foi avaliado pela medida do limiar de retirada da pata em ratos que receberam estimulações mecânica e térmica. Resultados: A estimulação diária com eletroacupuntura aumentou efetivamente o limiar de retirada da pata em ratos com Adjuvante Completo de Freund. A eletroacupuntura aumentou o nível de adenosina na região zusanli. Estudos posteriores mostraram que a eletroacupuntura poderia diminuir os níveis de substância P, receptor de neurocinina-1, fator de necrose tumoral-alpha, interleucina-1β, interleucina-6 e CD68 nos gânglios da raiz dorsal. Curiosamente, a injeção direta de antagonistas do receptor de adenosina A1 ou de substância P, ou a transecção da raiz do nervo dorsal, podem prejudicar significativamente as ações analgésicas induzidas pela eletroacupuntura em ratos com Adjuvante Completo de Freund e reduzir os níveis de substância P, receptor de neurocinina-1, fator de necrose tumoral-alfa, interleucina-1β, interleucina-6 e CD68. Por fim, confirmamos que a injeção direta de um agonista do receptor da adenosina A1 reproduziu os efeitos analgésicos da eletroacupuntura. Conclusão: Nossos resultados indicam a regulação da secreção da substância P mediada pela adenosina. A via mediada pela substância P pode estar envolvida no processo de analgesia por eletroacupuntura em ratos.

ABSTRACT Background: Acupuncture has been widely used for alleviating pain. However, its mechanisms remain largely enigmatic. Objective: In the present study, we focused on whether the analgesic effect of electroacupuncture is related to its regulation on adenosine and substance P expression. Methods: We established chronic inflammatory pain model in rats through a single injection of Complete Freund’s Adjuvant, and then we treated animals using daily electroacupuncture. We applied seven bilateral sessions of electroacupuncture (ST36 and BL60, 0.5 to 1.5 mA, initial strength of 0.5 mA, increased by 0.5 mA every 10 minutes, for 30 minutes per session, one section per day) to Complete Freund’s Adjuvant rats for seven days. The analgesic effect of electroacupuncture was evaluated by measuring paw withdrawal threshold in rats that received mechanical and thermal stimulation. Results: Daily electroacupuncture stimulation effectively increased paw withdrawal threshold in Complete Freund’s Adjuvant rats. Electroacupuncture increased the adenosine level in zusanli. A further study showed that electroacupuncture could decrease substance P, neurokinin-1 receptor, tumor necrosis factor-alpha, interleukin-1β, interleukin-6 and CD68 levels in dorsal root ganglion. Interestingly, direct injection of adenosine A1 or substance P receptor antagonists, or dorsal nerve root transection could significantly impair electroacupuncture induced analgesic actions in Complete Freund’s Adjuvant rats could and reduce the levels of substance P, neurokinin-1 receptor, tumor necrosis factor-alpha, interleukin-1β, interleukin-6 and CD68. Finally, we confirmed that direct injection of adenosine A1 receptor agonist replicated the analgesic effect of electroacupuncture. Conclusion: Our results indicate regulation of adenosine-mediated substance P secretion. Substance P-mediated pathway may be involved in the analgesia process by electroacupuncture in rats.

Abstract Background: Acupuncture, a therapy of traditional Chinese medicine, is confirmed to exert the therapeutic action on polycystic ovary syndrome (PCOS). However, the detailed therapeutic mechanisms of acupuncture in PCOS remain ambiguous. In this study, we further investigated whether electroacupuncture (EA) alleviated PCOS-like symptoms in rats via regulating a metabolic regulator, sterol regulatory element binding protein-1 (SREBP1). Methods: The PCOS-like rat model was built by hypodermic injection with dehydroepiandrosterone (DHEA). The rats were subjected to EA intervention (ST29 and SP6 acupuncture points) for 5 weeks. Primary granulosa cells were isolated from control and PCOS-like rats for evaluating insulin resistance, mitochondrial dysfunction and oxidative stress in vitro. Results: The expression of SREBP1 was increased in PCOS-like rats, which was suppressed by EA treatment. In addition, lentivirus-mediated overexpression of SREBP1 restrained EA treatment-induced improvement in pathological changes, serum hormone levels and insulin resistance in rats. In addition, overexpression of SREBP1 repressed insulin-stimulated phosphorylation of insulin receptor β (IR) and AKT in primary granulosa cells. Moreover, upregulation of SREBP1 further exacerbated mitochondrial dysfunction and oxidative stress in granulosa cells isolated from PCOS-like rats. Mechanically, EA treatment suppressed SREBP1 expression through inducing the activation of AMP-activated protein kinase (AMPK) signaling pathway in PCOS-like rats. Conclusion: EA intervention alleviated PCOS-like symptoms in rats via improving IR, mitochondrial dysfunction and oxidative stress through regulating SREBP1, a lipid metabolism regulator. Our findings illuminate the novel protective mechanisms of EA in the treatment of PCOS.

Abstract Purpose To investigate whether heat shock protein 90 (HSP90) is involved in complement regulation in ischemic postconditioning (IPC). Methods The left coronary artery of rats underwent 30 min of occlusion, followed by 120 min of reperfusion and treatment with IPC via 3 cycles of 30s reperfusion and 30s occlusion. The rats were injected intraperitoneally with 1 mg/kg HSP90 inhibitor geldanamycin (GA) after anesthesia. Eighty rats were randomly divided into four groups: sham, ischemia-reperfusion (I/R), IPC and IPC + GA. Myocardial infarct size, apoptosis index and the expression of HSP90, C3, C5a, tumor necrosis factor (TNF)-alpha, interleukin (IL)-1β and c-Jun N-terminal kinase (JNK) were assessed. Results Compared with the I/R injury, the IPC treatment significantly reduced infarct size, release of troponin T, creatine kinase-MB, and lactate dehydrogenase, and cardiomyocyte apoptosis. These beneficial effects were accompanied by a decrease in TNF-α, IL-1β, C3, C5a and JNK expression levels. However, all these effects were abrogated by administration of the HSP90 inhibitor GA. Conclusion HSP90 exerts a profound effect on IPC cardioprotection, and may be linked to the inhibition of the complement system and JNK, ultimately attenuating I/R-induced myocardial injury and apoptosis.

SUMMARY: The experiment was conducted to evaluate the effects of dietary supplemental chromium (Cr) on growth performance, meat quality, intestinal morphology, mucosa Hsp70 mRNA expression and antioxidant status of ducks reared under heat stress conditions. All ducks were randomly divided into three treatment groups, respectively, control group (Control, 23 ± 2 °C), heat stress group (HS, 32 ±2 °C), Cr picolinate group (CrPic, 32 ± 2 °C, 0.2 mg Cr/kg). Feed and distilled-deionized water were available ad libitum for an experimental phase of 35 days. Samples were collected on the day 14, 21 and 35 to determine biological and hematological values. Results showed that heat stress or dietary supplemental Cr both didn’t have distinct influence on growth performance (P>0.05), compared to controls. Ducks fed 0.2 mg Cr/kg diet had greater ultimate pH (pHu)(P<0.05) than HS group. At day 14, the ratio of villus height to crypt depth (V/C) in CrPic group significantly increased (P<0.05) than that of HS group in jejunum. Heat stress remarkably increased Hsp70 mRNA expression in jejunum compared with controls (P<0.05). While the expression of Hsp70 mRNA in CrPic group was significantly decreased compared with HS (P<0.05). At day 21, the V/C of ileum in CrPic group significantly increased compared with HS group (P<0.05). Serum SOD levels in CrPic group were significantly higher than those in HS group (P<0.05). At day 35, Hsp70 mRNA expression and serum T-SOD levels in CrPic group significantly increased compared with controls (P<0.05). T-AOC in HS group significantly decreased compared with controls (P<0.05). Results indicate that dietary Cr supplementation doesn’t influence ducks’ growth performance, but has a positive effect on meat quality, small intestine morphology, also regulates Hsp70 mRNA expression under heat stress conditions, and enhances the antioxidant status.

RESUMEN: Se evaluó los efectos del cromo (Cr) dietético suplementario sobre el rendimiento del crecimiento, la calidad de la carne, la morfología intestinal, la expresión del ARNm Hsp70 en la mucosa y el estado antioxidante de los patos criados bajo condiciones de estrés por calor. Todos los patos se dividieron aleatoriamente en tres grupos: grupo control (control, 23 ± 2 °C), grupo de estrés térmico (HS, 32 ± 2 °C) y grupo de picolinato de Cr (CrPic, 32 ± 2 °C, 0,2 mg Cr / kg). El alimento y el agua desionizada destilada estuvieron disponibles ad libitum durante la fase experimental de 35 días. Las muestras se recogieron los días 14, 21 y 35 para determinar los valores biológicos y hematológicos. Los resultados mostraron que el estrés térmico o la suplementación dietética de Cr no tuvieron una influencia distinta en el rendimiento del crecimiento (P> 0,05), en comparación con los controles. Los patos alimentados con 0,2 mg de Cr / kg de dieta tuvieron un mayor pH final (pHu) (P <0,05) que el grupo HS. En el día 14, la relación de la altura de las vellosidades a la profundidad de la cripta (V / C) en el grupo CrPic aumentó significativamente (P <0,05) en relación a la del grupo de HS en el yeyuno. El estrés por calor incrementó notablemente la expresión del ARNm de Hsp70 en el yeyuno en comparación con los controles (P <0,05). Mientras que la expresión del ARNm de Hsp70 en el grupo CrPic se redujo significativamente en comparación con HS (P <0,05). En el día 21, la relación V / C del íleon en el grupo CrPic aumentó significativamente en comparación con el grupo HS (p <0,05). Los niveles séricos de SOD en el grupo CrPic fueron significativamente más altos que los del grupo HS (P <0,05). En el día 35, la expresión de ARNm de Hsp70 y los niveles séricos de T-SOD en el grupo CrPic aumentaron significativamente en comparación con los controles (P <0,05). T-AOC en el grupo HS disminuyó significativamente en comparación con los controles (P <0,05). Los resultados indican que la suplementación dietética de Cr no influye en el rendimiento de crecimiento de los patos, pero tiene un efecto positivo en la calidad de la carne, en la morfología del intestino delgado, y también regula la expresión de ARNm de Hsp70 en condiciones de estrés calórico y mejora el estado antioxidante.

Abstract Background: SET domain bifurcated 1 (SETDB1) has been widely considered as an oncogene playing a critical role in many human cancers, including breast cancer. Nevertheless, the molecular mechanism by which SETDB1 regulates breast cancer tumorigenesis is still unknown. Methods: qRT-PCR assay or western blot analysis was performed to assess the expression level of SETDB1 mRNA or protein, respectively. siSETDB1, pCMV6-XL5-SETDB1, miR-381-3p mimic, or miR-381-3p inhibitor was transfected into cells to regulate the expression of SETDB1 or miR-381-3p. MiRNA directly interacted with SETDB1 was verified by luciferase reporter assay and RNA immunoprecipitation. CCK-8 assay, colony formation assay, flow cytometric analysis, and transwell assay were used to detect the abilities of cell proliferation, cell cycle progression and migration, respectively. Animal model of xenograft tumor was used to observe the regulatory effect of SETDB1 on tumor growth in vivo. Results: We verified that SETDB1 mRNA level was upregulated in breast cancer tissues and cell lines, and SETDB1 depletion led to a suppression of cell proliferation, cell cycle progression and migration in vitro, as well as tumor growth in vivo. SETDB1 was verified to be a target of miR-381-3p. Moreover, miR-381-3p overexpression suppressed cell proliferation, cell cycle progression and migration, whereas SETDB1 abated miR-381-3p-mediated regulatory function on breast cancer cells. Conclusions: This study revealed that SETDB1 knockdown might suppress breast cancer progression at least partly by miR-381-3p-related regulation, providing a novel prospect in breast cancer therapy.

RESUMEN Para estimar la colapsabilidad en Xixian se realizó el test de compresión (K o ), de acuerdo con el mecanismo de colapsabilidad, con diferentes combinaciones de presión (0~1.2 MPa) y contenido de agua (4%~Sat). Basado en la curva σ-ε con diferentes contenidos de agua se calcularon los coeficientes de asentamiento y colapsabilidad a partir de substracción de la curva en condiciones de saturación y analizando los efectos de presión y contenido de agua. Los resultados muestran que el promedio de colapsabilidad por su propio peso para el área de Xixian es de nivel II, con un límite bajo de hasta 14 m. La compresibilidad de loess es proporcional a la presión y al contenido de agua. Con un bajo contenido de agua, el coeficiente de colapsabilidad (δ s ) se incrementó mientras la presión se aumentó; pero, con contenidos medios y altos de agua, el coeficiente de colapsabilidad alcanzó su pico con el incremento de la presión, y luego bajó para prácticamente estabilizarse. Bajo presión constante, la colapsabilidad descendió al aumentar el contenido de agua. Si se asume como criterio inicial de colapso (P i ) una deformación adicional de 1.5%, Pi es linealmente proporcional al contenido de agua. El contenido de agua inicial de colapso (w i ) se incrementa drásticamente cuando la presión se eleva a baja presión, sin embargo, el contenido de agua alcanza un valor constante de 26 °% cuando la presión es de más de 200 kPa. Estos resultados son significativos para la investigación geotécnica y el diseño estructural en Xixian.

ABSTRACT More than 40% area of the Xixian New Area is a loess deposit region, and most of the loess landform is tableland and terrace where the thickness of loess is considerable large. Therefore, loess collapsibility will be the most important geotechnical problem in future foundation investigation and construction. To explore loess collapsibility in the Xixian New Area, we conducted the K 0 compression test, based on the collapsibility mechanism, which has different combinations of pressure (0~1.2 MPa) and water content (4%~Sat). Based on the σ-ε curve under different water content, we calculated the generalized collapse settlement and collapsibility coefficient of every water content under every pressure by subtracting the relevant curve from the saturated curve and analyzed the cross action of pressure and water on loess compressibility. The results show that the average collapsibility level of the northern Xixian New Area is self-weight collapsible level II, with a lower limit of 14 m. The compressibility of loess is proportional to pressure and water content. Under low water content, the collapsibility coefficient δ s increased while the pressure increased, but under medium and high water content, δ s will reach a peak with increasing pressure and after that, δ s will decrease until its value is close to constant. When under the same pressure, δ s decreases when water content increases. If set the additional strain 1.5% as collapse start criterion, then the first collapse pressure P i will linear proportional to water content. The initial collapse water content w i will increase sharply when pressure raises under low pressure, but w i will reach a constant value of 26% when pressure is more than 200 kPa. This consequence will be meaningful for future geotechnical investigation and design in the Xixian New Area.

Abstract: Introduction and aim. Hyponatremia is common in patients with decompensated cirrhosis and is associated with increased mortality. Tolvaptan, a vasopressor V2 receptor antagonist, can increase free wáter excretion, but its efficacy and safety in cirrhotic patients remain unclear. Material and methods. We studied the usage and safety of tolvaptan in cirrhotic patients in a real-life, non-randomized, multicenter prospective cohort study. Forty-nine cirrhotic patients with hyponatremia were treated with tolvaptan 15 mg daily, and 48 patients not treated with tolvaptan in the same period served as controls. Improvement in serum sodium level was defined as an increase in serum sodium from < 125 to ≥ 125 mmol/L or from 125-134 to ≥ 135 mmol/L on day 7. Results. Twenty-three (47%) patients in the tolvaptan group and 17 (35%) in the control group had normal serum sodium on day 7 (p = 0.25). Serum sodium improved in 30 (61%) patients in the tolvaptan group and 17 (35%) patients in the control group (p = 0.011). Adverse events occurred in 46-47% of patients in both groups, and tolvaptan was not associated with worsened liver function. No patient with normal serum sodium on day 7 died within 30 days of treatment, whereas 16% of those with persistent hyponatremia died (p = 0.0019). Conclusion. In conclusion, short-term tolvaptan treatment is safe and can improve serum sodium level in cirrhotic patients with hyponatremia. Normalization of serum sodium level is associated with better survival.

ABSTRACT Safflower (Carthamus tinctorius L., Asteraceae) is an important oil crop and medicinal plant. Gene expression analysis is gaining importance in the research of safflower. Quantitative PCR has become a powerful method for gene study. Reference genes are one of the major qualification requirements of qPCR because they can reduce the variability. To identify the reference genes in safflower, nine candidate genes of the housekeeping genes were selected from the EST library of safflower constructed by our lab: CtACT (actin), CtGAPDH (glyceraldehyde 3-phosphate dehydrogenase), CtE1F4A (elongation factor 1 alpha), CtTUA (alpha-tubulin), CtTUB (beta-tubulin), CtPP2A (serine/threonine-protein phosphatase), CtE1F4A (eukaryotic initiation factor 4A), CtUBI (Ubiquitin), and Ct60S (60S acidic ribosomal protein). Expression stability was examined by qPCR across 54 samples, representing tissues at different flowering stages and two chemotype of safflower lines. We assessed the expression stability of these candidate genes by employing four different algorithms (geNorm, NormFinder, ΔCt approach, and BestKeeper) and found that CtUBI and Ct60S were the highly ranked candidate genes. CtUBI and Ct60S were used as reference genes to evaluate the expression of CtFAD2-10 and CtKASII. Our data suggest CtUBI and Ct60S could be used as internal controls to normalize gene expression in safflower.

Prompted by the 20th anniversary of the 1993 World Development Report, a Lancet Commission revisited the case for investment in health and developed a new investment framework to achieve dramatic health gains by 2035. The Commission's report has four key messages, each accompanied by opportunities for action by national governments of low-income and middle-income countries and by the international community. First, there is an enormous economic payoff from investing in health. The impressive returns make a strong case for both increased domestic financing of health and for allocating a higher proportion of official development assistance to development of health. Second, modeling by the Commission found that a "grand convergence" in health is achievable by 2035-that is, a reduction in infectious, maternal, and child mortality down to universally low levels. Convergence would require aggressive scale up of existing and new health tools, and it could mostly be financed from the expected economic growth of low- and middle-income countries. The international community can best support convergence by funding the development and delivery of new health technologies and by curbing antibiotic resistance. Third, fiscal policies -such as taxation of tobacco and alcohol- are a powerful and underused lever that governments can use to curb non-communicable diseases and injuries while also raising revenue for health. International action on NCDs and injuries should focus on providing technical assistance on fiscal policies, regional cooperation on tobacco, and funding policy and implementation research on scaling-up of interventions to tackle these conditions. Fourth, progressive universalism, a pathway to universal health coverage (UHC) that includes the poor from the outset, is an efficient way to achieve health and financial risk protection. For national governments, progressive universalism would yield high health gains per dollar spent and poor people would gain the most in terms of health and financial protection. The international community can best support countries to implement progressive UHC by financing policy and implementation research, such as on the mechanics of designing and implementing evolution of the benefits package as the resource envelope for public finance grows.

Con motivo del 20º aniversario del Informe sobre el Desarrollo Mundial 1993, una Comisión de la revista The Lancet reconsideró el argumento a favor de la inversión en salud y desarrolló un nuevo marco de inversión para lograr mejoras dramáticas en materia de salud para el año 2035. El informe de la Comisión contiene cuatro mensajes clave, cada uno acompañado de oportunidades para los gobiernos nacionales de países de ingresos bajos y medios y para la comunidad internacional. En primer lugar, invertir en salud acarrea enormes rendimientos económicos. Las impresionantes ganancias son un fuerte argumento a favor de un aumento en el financiamiento nacional de la salud y de asignar una mayor proporción de la asistencia oficial al desarrollo de la salud. En segundo lugar, en el modelo creado por la Comisión se encontró que es posible lograr para el año 2035 una "gran convergencia" en salud, consistente en la reducción de las tasas de mortalidad materna, infantil y por infecciones a niveles universalmente bajos. Tal convergencia requeriría la ampliación de las herramientas de salud existentes y un incremento agresivo de nuevas herramientas, y podría ser financiada en su mayor parte con recursos derivados del crecimiento económico esperado de los países de ingresos bajos y medios. La mejor manera en que la comunidad internacional puede apoyar la convergencia es financiando el desarrollo y suministro de nuevas tecnologías de salud, y frenando la resistencia a los antibióticos. En tercer lugar, las políticas fiscales -tales como los impuestos al tabaco y al alcohol- son una palanca poderosa y subutilizada que los gobiernos pueden emplear para detener el avance de las enfermedades no transmisibles (ENT) y las lesiones, a la vez que elevan los ingresos públicos para la salud. La acción internacional sobre las ENT y lesiones debería enfocarse en proporcionar asistencia técnica sobre políticas fiscales, en cooperación regional para el combate al tabaquismo y en financiar investigación sobre políticas e implementación para ampliar las intervenciones que enfrenten estos problemas. En cuarto lugar, la universalización progresiva -una vía hacia la cobertura universal de salud (CUS) que incluya desde el comienzo a los pobres- es una manera eficiente de lograr la protección a la salud contra riesgos financieros. Para los gobiernos nacionales, la universalización progresiva produciría elevadas ganancias en salud por cada dólar que se gaste en ésta, y los pobres serían quienes más ganarían en términos tanto de salud como de protección financiera. La mejor manera en que la comunidad internacional puede brindar apoyo a los países para implementar una CUS progresiva es financiando la investigación sobre políticas e implementación, por ejemplo, sobre la mecánica del diseño e instrumentación de la evolución del paquete de beneficios conforme crezca el presupuesto para las finanzas públicas.