ABSTRACT Blue eggshells have become economically valuable due to their higher popularity among consumers when compared with eggs of other colors, particularly in East Asia. However, eggshell colors vary widely and gradually become lighter with the increase of age. Therefore, to determine the association between DNA methylation and gene expression on the molecular mechanism of eggshell color variation in Xuefeng Black-bone chickens, we collected the shell glands from chickens that produced dark blue eggshells and light blue eggshells, respectively, and performed whole genome bisulfite sequencing (WGBS) and RNA sequencing (RNA-Seq) on them. The results showed that in the context of CG, DNA methylation levels were the highest compared to CHG and CHH contexts, and were negatively correlated with gene expression levels in three different gene regions in all samples, while there was no significant correlation in the contexts of CHG and CHH. Furthermore, when disregarding the location of coding genes, a total of 55 genes will show differences not only in expression levels but also in DNA methylation levels, and among the majority of differential expression genes (n=50), the DNA methylation levels in the gene body region were the highest. GO analysis and KEGG pathway analysis were performed in these genes and the results showed that the genes NR1H4 and HEPHL1, ABCA4 and ABCA12, and the pathways, cell adhesion molecular, ABC transporters, ECM-receptor interaction, and neuroactive ligand-receptor interaction, may affect the color variation of the blue eggshell of Xuefeng Black-bone chickens.

The global burden of chronic kidney disease (CKD) is rapidly increasing with a projection of becoming the 5th most common cause of years of life lost globally by 2040. CKD is a major cause of catastrophic health expenditure. The costs of dialysis and transplantation consume up to 3% of the annual healthcare budget in high-income countries. However, the onset and progression of CKD is often preventable. In 2020, the World Kidney Day campaign highlights the importance of preventive interventions – be it primary, secondary, or tertiary. This article focuses on outlining and analyzing measures that can be implemented in every country to promote and advance CKD prevention. Primary prevention of kidney disease should focus on the modification of risk factors and addressing structural abnormalities of the kidney and urinary tracts, as well as exposure to environmental risk factors and nephrotoxins. In persons with pre-existing kidney disease, secondary prevention, including blood pressure optimization and glycemic control, should be the main goal of education and clinical interventions. In patients with advanced CKD, management of co-morbidities such as uremia and cardiovascular disease is a highly recommended preventative intervention to avoid or delay dialysis or kidney transplantation. Political efforts are needed to proliferate the preventive approach. While national policies and strategies for non-communicable diseases might be present in a country, specific policies directed toward education and awareness about CKD screening, management, and treatment are often lacking. Hence, there is an urgent need to increase the awareness of preventive measures throughout populations, professionals, and policy makers.

Abstract The influence of water resources allocation patterns on root layers soil desalination and groundwater quality were investigated by monitoring different irrigation area from People’s Victory Canal irrigation district in Huang Huai Hai plain China that had been conducted well canal combined irrigation patterns from 1954. A typical area was chosen belong to the People’s Victory Canal irrigation district from 2013 to 2015. Precipitation of the area, surface water irrigation amount, groundwater consumption amount, salinity content in 0 to 100 cm soil layer and total dissolved solids in groundwater were monitored, the ratio of canal-well water (the ratio of surface water to groundwater irrigation amount, CWWR), soil salinity spatial dynamic, desalinization rate in 0 to 100 cm soil layer and sodium adsorption ratio (SAR) of groundwater were analyzed in the area. CWWR of Branch 1, Branch 2, and Branch 3 ranged from 0.72 to 1.03, 2.50 to 2.63, and 0.65 to 1.26 in 2013 to 2015, respectively. The soil salinity contents for the top 100 cm horizons in the Branch 2 irrigation area decreased slightly by 1.63% to 8.90% compared with the values in the Branch 1 and Branch 3 irrigation districts while the water resources allocation patterns were conducted two years later. Compared with the area in the hydrological period in 2013, the area of average soil salinity exceeded 0.32 mS/cm for the top 20 cm horizons has decreased notably in 2014 and 2015. Compared with the values in 2014, the average TDS concentration of groundwater increased by 18.66% in the study area in 2015, whereas the sodium adsorption ratio (SAR) values in the above-mentioned irrigation areas decreased by 23.58%, 36.82%, and 53.37% in the normal period in 2015. The combined well-canal irrigation pattern could represent a useful water allocation method mainly because of the temporal and spatial regulation of water resources and root layer soil desalination; however, possible increases of TDS in the groundwater may cause potential ecosystem degradation risks after long-term water allocation in semi-arid areas. Thus, water allocation should be emphasized to maintain a healthy groundwater environment and sustainable stable yields of grain in combined canal-well irrigation districts.

MicroRNAs (miRNAs) play an important role in drug resistance and modulate the efficiency of chemotherapy. A recent study indicated that miR-340 functions as a tumor suppressor in various types of cancer. However, the role of miR-340 in chemotherapy has not been reported yet. In this study, we found that miR-340 enhanced cisplatin (CDDP)-induced cell death. Induction of miR-340-5p expression decreased the IC50 of CDDP and increased the apoptosis of CDDP-resistant MG-63 and Saos-2 cells. Moreover, miR-340-5p decreased the accumulation of MRP1 and MDR1. We further explored the mechanism underlying the promoting effects of miR-340-5p on CDDP-induced cell death. We identified a potential target of miR-340 in the 3′ untranslated region of lysophosphatidic acid acyltransferase (LPAATβ) using the online program Targetscan (http://www.microrna.org). Luciferase reporter assays showed that miR-340 binds to the 3′UTR of LPAATβ. Enforced expression of miR-340-5p decreased the accumulation of LPAATβ in both MG-63 and Saos-2 cells. Silencing LPAATβ decreased the IC50 of CDDP and increased the apoptosis of CDDP-resistant MG-63 and Saos-2 cells, which is consistent with the effect of miR-340-5p on CDDP-induced cell death. Moreover, induced expression of LPAATβ compromised the effects of miR-340-5p on CDDP-induced cell death and accumulation of MRP1 and MDR1. Taken together, our data indicated that miR-340-5p enhanced the sensitivity to CDDP by targeting LPAATβ.

Glycyrrhizin has been used clinically for several years due to its beneficial effect on immunoglobulin E (IgE)-induced allergic diseases, alopecia areata and psoriasis. In this study, glycyrrhizin, ultraviolet B light (UVB) or a combination of both were used to treat active-stage generalized vitiligo. One hundred and forty-four patients between the ages of 3 and 48 years were divided into three groups: group A received oral compound glycyrrhizin (OCG); group B received UVB applications twice weekly, and group C received OCG+UVB. Follow-ups were performed at 2, 4, and 6 months after the treatment was initiated. The Vitiligo Area Scoring Index (VASI) and the Vitiligo Disease Activity (VIDA) instrument were used to assess the affected body surface, at each follow-up. Results showed that 77.1, 75.0 and 87.5% in groups A, B and C, respectively, presented repigmentation of lesions. Responsiveness to therapy seemed to be associated with lesion location and patient compliance. Adverse events were limited and transient. This study showed that, although the three treatment protocols had positive results, OCG and UVB combination therapy was the most effective and led to improvement in disease stage from active to stable.

Affective states influence subsequent attention allocation. We evaluated emotional negativity bias modulation by reappraisal in patients with generalized anxiety disorder (GAD) relative to normal controls. Event-related potential (ERP) recordings were obtained, and changes in P200 and P300 amplitudes in response to negative or neutral words were noted after decreasing negative emotion or establishing a neutral condition. We found that in GAD patients only, the mean P200 amplitude after negative word presentation was much higher than after the presentation of neutral words. In normal controls, after downregulation of negative emotion, the mean P300 amplitude in response to negative words was much lower than after neutral words, and this was significant in both the left and right regions. In GAD patients, the negative bias remained prominent and was not affected by reappraisal at the early stage. Reappraisal was observed to have a lateralized effect at the late stage.

ABSTRACT Objective: This study investigates the change of endothelial cell morphology and function at the rabbit basilar bifurcations in response to sustained high blood flow after bilateral common carotid artery ligation. Methods: Fifteen adult female New Zealand white rabbits were divided into experimental and sham control groups. The experimental group was subjected to bilateral common carotid artery ligation to increase the compensatory basilar artery flow. Basilar artery flow was monitored by transcranial Doppler after surgery. The endothelial cells at the arterial bifurcations were studied morphologically by electron microscopy and immunohistochemistry using β-catenin antibodies. Basilar artery flow increased significantly following common carotid artery ligation. Results: Early-stage basilar artery bifurcation aneurysms were present in all rabbits at three months after ligation. The endothelial cells changed from a fusiform to column shape at the basilar artery bifurcation. Gaps between endothelial cells of the experimental group appeared wider in the electron microscopic photographs compared with those of the control group. The expression of endothelial β- catenin at the arterial bifurcations also decreased. Conclusion: This study is the first to present endothelial cell changes of basilar artery bifurcation in response to sustained high blood flow in rabbits. Endothelial cell impairment possibly initiates aneurysm formation.

RESUMEN Objetivo: Este estudio investiga el cambio de morfología de la célula endotelial y la función en las bifurcaciones basilares del conejo en respuesta al flujo sanguíneo alto sostenido después de la ligadura de la arteria carótida común bilateral. Métodos: Quince conejos blancos adultos hembras de Nueva Zelanda fueron divididos en un grupo experimental y un grupo de control con procedimiento simulado. El grupo experimental fue sometido a la ligadura de la arteria carótida común bilateral para aumentar el flujo de la arteria basilar compensatoria. El flujo de la arteria basilar fue monitoreado mediante Doppler transcraneal después de la cirugía. Las células endoteliales en las bifurcaciones arteriales fueron estudiadas morfológicamente mediante microscopia electrónica e inmunohistoquímica usando anticuerpos β-catenina. El flujo de la arteria basilar aumentó considerablemente tras la ligadura de la arteria carótida común. Resultados: Aneurismas de la bifurcación de la arteria basilar producidos en etapa temprana, se hallaban presentes en todos los conejos tres meses después de la ligadura. Las células endoteliales cambiaron de forma fusiforme a forma de columna en la bifurcación de la arteria basilar. Los espacios entre las células endoteliales del grupo experimental aparecían con mayor amplitud en las fotografías microscópicas electrónicas comparados con los del grupo de control. También disminuyó la expresión de β-catenina endotelial en las bifurcaciones arteriales. Conclusión: Este estudio es el primero en presentar cambios de la célula endotelial de la bifurcación de la arteria basilar en respuesta al flujo sanguíneo alto sostenido en conejos. El deterioro de la célula endotelial posiblemente inicia la formación de aneurismas.

The objective of the present study was to explore the factors related to the prognosis of colorectal cancer (CRC) and to establish a prognostic model for the selection of patients who might benefit from hepatic resection for metastatic CRC. A total of 293 patients undergoing liver resection for metastatic CRC (172 males and 80 females ranging in age from 26 to 80 years) were selected and clinical, pathological and outcome data were examined in this retrospective study. The prognostic index (PI) of the patients was calculated on the basis of results of multivariate analysis. Patients were stratified into different groups, with survival curves projected according to PI. The 1-, 3-, and 5-year overall survival rates were 58.3, 26.4, and 11.3%, respectively. Univariate analysis indicated that degree of primary tumor differentiation, resection margin, preoperative carcinoembryonic antigen (CEA) level, number of liver metastases, and resection of liver metastases were associated with prognosis (P < 0.05). In multivariate analysis, the last three factors were found to be independent prognostic factors. The resection of liver metastases was a favorable factor. Patients were classified into three groups according to PI, which differed significantly in survival rate (P < 0.05). The individual survival rate was evaluated based on PI. Resection of hepatic colorectal metastases may produce long-term survival and cure. The proposed PI was easy to use, was highly predictive of patient outcome, and permitted categorization of patients into treatment groups.

Hepatocellular carcinoma (HCC) is one of the most common malignant tumours in the world, especially in Guangxi, China. The causes and mechanism of its tumourigenesis and development have not been completely clarified. Some studies revealed that the hepatic local cellular immune function was one of the factors. In the present study, the local micro-environmental immune status was explored by investigating the number, distribution and function of CD3, CD57,CD20, CD68, and granzyme B (GrB) positive cells in 60 patients with HCC and 62 patients with liver cirrhosis (LC) and its relationship with the prognosis of the patients. The results showed that the number of T and B lymphocytes and natural killer (NK) cells in the liver of HCC patients was significantly higher than that in the LC and normal controls; while the number of macrophages (Mj) was significantly lower. The number of Mj in the tissues decreased successively with the decrease of HCC differentiation; GrB-expressing cells in the liver predominantly consisted of CD57 positive cells. The number of NK cells, B lymphocytes and GrB-expressing cells in the cancerous tissues of stage I and II was significantly higher than that of stages III and IV. The number of T lymphocytes, NK cells, Mj and GrB-expressing lymphocytes in HCC cases without metastasis in 15 months was significantly higher than in the metastatic counterparts. The number of T and B lymphocytes, NK cells, and GrB-expressing cells decreased in patients with the progression of the HCC. These results suggest that the number of T and B lymphocytes, NK cells, Mj and GrB-positive lymphocytes might be important markers in the estimation of hepatic local immune status and be useful factors for predicting the prognosis of HCC patients.

El carcinoma hepatocelular (CHC) es uno de los tumores malignos más comunes en el mundo, especialmente en Guangxi, China. Las causas y el mecanismo de su desarrollo y génesis tumoral no han sido completamente aclarados. Algunos estudios revelaron que la función inmune celular local hepática era uno de los factores. En este estudio se exploró el estatus inmune microambiental local, mediante la investigación del número, distribución y función de CD3, CD57,CD20, CD68, y células positivas para la granzima B (GrB) en 60 pacientes con CHC y 62 pacientes con cirrosis hepática (CH) y su relación con la prognosis de los pacientes. Los resultados mostraron que el número de linfocitos T y B y las células asesinas naturales (NK) en el hígado de los pacientes con HCH, era significativamente mayor que en los controles normales y los pacientes con CH, mientras que el número de macrófagos (Mj) fue significativamente menor. El número de Mj en los tejidos disminuyó sucesivamente a la par con la disminución de la diferenciación de CHC; las células expresoras de GrB en el hígado consistían predominantemente en células positivas CD57. El número de células NK, linfocitos B y células expresoras de GrB en los tejidos cancerosos de etapas I y II, fue significativamente más alto que el de las etapas III y IV. El número de linfocitos T, células NK, Mj y linfocitos expresores de GrB en los casos de CHC sin metástasis en 15 meses, fue significativamente mayor que en las contrapartidas metastáticas. El número de linfocitos T y B, células NK, y células expresoras de GrB, disminuyó en los pacientes a medida que progresaba el CHC. Estos resultados sugieren que el número de linfocitos T y B, células NK, Mj y linfocitos GrB-positivos podrían ser marcadores importantes en la valoración del estatus inmune local hepático y los factores útiles a la hora de predecir la prognosis de los pacientes de CHC.

Objectives: To determine cytotoxic activity of fractions from a 1:1 combination of an ethanol extract of Annona muricata leaves (soursop) and atomized aqueous extract of Krameria lappacea root (Ratania) in breast (MCF-7), lung (H-460), and central nervous system (SF-268) cancer cell cultures. Material and methods: For fractionating the 1:1 mixture of Annona and Krameria a 50-cm long chromatographic column was prepared using dichloromethane, dichloromethane: ethyl acetate and ChCl 3:MeOH as increasing polarity eluting systems and 186 fractions were obtained. Fractions 2 to 83 were assessed in breast (MCF-7), lung (H-460), and central nervous system (SF-268) cancer cell cultures. All fractions were assessed two times. Those fractions that showed <50% growth of cancer cells (%G) in any of the three cell lines were assayed once again using five different concentrations, in order to determine the concentration where there was a 50% inhibition of cancer cell growth (GI 50 ). Those fractions with a <10 m g/mL GI 50 were considered to be active against cancer cell lines. Results: Fractions 7 to 17 of the association of the two aforementioned natural products has GI 50 values reported as 1,6, 1,4, and 1,4 m g/mL against MCF-7, H-460, and SF-268 cancer cell lines, respectively. Conclusions: Fractions 7 to 17 of the Annona and Krameria combination showed in vitro cytotoxic activity against breast, lung, and central nervous system cancer cultured cells.

Objetivos: Determinar la actividad citotóxica de las fracciones procedentes de la combinación 1:1 del extracto etanólico de hojas de Annona muricata L (guanábana) y el extracto acuoso atomizado de la raíz de Krameria lappacea (ratania) en cultivos de líneas celulares cancerosas de glándula mamaria (MCF-7), pulmón (H-460) y Sistema nervioso central (SF-268. Materiales y métodos: Para el fraccionamiento de la mezcla 1:1 de Annona mas Krameria se preparó una columna cromatográfica de 50 cm de longitud empleando diclorometano, diclorometano: acetato de etilo y CHCl 3:MeOH como sistemas de elusión de polaridad creciente, obteniéndose 186 fracciones. Se evaluaron las fracciones 2 a 83 en cultivo de células cancerosas de glándula mamaria (MCF-7), de pulmón (H-460) y del sistema Nervioso central (SF-268. Todas las fracciones fueron ensayadas en duplicado. Aquellas fracciones que presenta-ron un porcentaje de crecimiento de células cancerosas (%G) <50% en alguna de las tres líneas celulares fueron ensayadas nuevamente a cinco concentraciones, para determinar finalmente la concentración a la cual se inhibe el 50% del crecimiento de las células cancerosas (GI 50. Se consideraron activas aquellas fracciones con una GI 50 <10 µg/mL. Resultados: Las fracciones 7 a 17 procedentes de la asociación de los dos productos naturales frente a los cultivos de las líneas celulares tumorales MCF-7, H-460 y SF-268 mostraron una GI 50 de 1,6, 1,4 y 1,4 µg/mL respectivamente. Conclusiones: Las fracciones 7 a 17 procedentes de la asociación de Annona más Krameria mostraron acción citotóxica in vitro frente al cultivo de células cancerosas de glándula mamaria, pulmón y del sistema nervioso central.

Estrogen stimulates the renin-angiotensin system by augmenting both tissue and circulating levels of angiotensinogen and renin. We show, however, that angiotensin converting enzyme (ACE) activity in the circulation and in tissues is reduced in two animal models of postmenopausal chronic hormone replacement. We observed a reduction of ACE activity in association with a significant increase in plasma angiotensin I (Ang I) and hyperreninemia in ovariectomized monkeys treated with Premarin (conjugated equine estrogen) replacement for 30 months. Plasma angiotensin II (Ang II) levels were not increased in monkeys treated with estrogen, suggesting that the decrease in ACE curtailed the formation of the peptide. The Ang II/Ang I ratio, an in vivo index of ACE activity, was significantly reduced by estrogen treatment, further supporting the biochemical significance of estrogen's inhibition of ACE. In ovariectomized transgenic hypertensive (mRen2)27 rats submitted to estrogen replacement treatment for 3 weeks, ACE activity in plasma and tissue (aorta and kidney) and circulating Ang II levels were reduced, whereas circulating levels of angiotensin-(1-7) (Ang-(1-7) were increased. Ang-(1-7), the N-terminal fragment of Ang II, is a novel vasodilator and antihypertensive peptide. Thus, the net balance of these effects of estrogen on the renin-angiotensin vasoconstrictor/vasodilator system is to promote the antihypertensive effect.