Guijing2501 (Citrus unshiu) Has Stronger Cold Tolerance Due to Higher Photoprotective Capacity as Revealed by Comparative Transcriptomic and Physiological Analysis and Overexpression of Early Light-Induced Protein.

Cold is one of the major limiting factors for citrus production, particularly extreme cold waves. Therefore, it is of great importance to develop cold-tolerant varieties and clarify their cold tolerance mechanisms in citrus breeding. In this study, comparative transcriptomic and physiological analyses were performed to dissect the cold tolerance mechanism of Guijing2501 (GJ2501), a new satsuma mandarin (Citrus unshiu) variety with about 1 °C lower LT50 (the median lethal temperature) relative to Guijing (GJ). The physiological analysis results revealed that GJ2501 is more cold-tolerant with less photoinhibition, PSII photodamage, and MDA accumulation, but higher POD activity than GJ under cold stress. Comparative transcriptomic analysis identified 4200 DEGs between GJ and GJ2501, as well as 4884 and 5580 up-regulated DEGs, and 5288 and 5862 down-regulated DEGs in response to cold stress in GJ and GJ2501, respectively. "Photosynthesis, light harvesting" and "photosystem" were the specific and most significantly enriched GO terms in GJ2501 in response to cold stress. Two CuELIP1 genes (encoding early light-induced proteins) related to the elimination of PSII photodamage and photoinhibition were remarkably up-regulated (by about 1000-fold) by cold stress in GJ2501 as indicated by RT-qPCR verification. Overexpression of CuELIP1 from GJ2501 in transgenic Arabidopsis protected PSII against photoinhibition under cold stress. Taken together, the cold tolerance of GJ2501 may be ascribed to its higher photoprotective capacity under cold stress.

A Novel Non-Specific Lipid Transfer Protein Gene, CmnsLTP6.9, Enhanced Osmotic and Drought Tolerance by Regulating ROS Scavenging and Remodeling Lipid Profiles in Chinese Chestnut (Castanea mollissima Blume).

Chestnut (Castanea mollissima Blume) is an important economic tree owing to its tasty fruit and adaptability to environmental stresses, especially drought. Currently, there is limited information about non-specific lipid transfer protein (nsLTP) genes that respond to abiotic stress in chestnuts. Here, a chestnut nsLTP, named CmnsLTP6.9, was identified and analyzed. The results showed that the CmnsLTP6.9 protein localized in the extracellular matrix had two splicing variants (CmnsLTP6.9L and CmnsLTP6.9S). Compared with CmnsLTP6.9L, CmnsLTP6.9S had an 87 bp deletion in the 5'-terminal. Overexpression of CmnsLTP6.9L in Arabidopsis enhanced tolerance to osmotic and drought stress. Upon exposure to osmotic and drought treatment, CmnsLTP6.9L could increase reactive oxygen species (ROS)-scavenging enzyme activity, alleviating ROS damage. However, CmnsLTP6.9S-overexpressing lines showed no significant differences in phenotype, ROS content, and related enzyme activities compared with the wild type (WT) under osmotic and drought treatment. Moreover, lipid metabolism analysis confirmed that, unlike CmnsLTP6.9S, CmnsLTP6.9L mainly altered and upregulated many fatty acyls and glycerophospholipids, which implied that CmnsLTP6.9L and CmnsLTP6.9S played different roles in lipid transference in the chestnut. Taken together, we analyzed the functions of CmnsLTP6.9L and CmnsLTP6.9S, and demonstrated that CmnsLTP6.9L enhanced drought and osmotic stress tolerance through ROS scavenging and lipid metabolism.

Evolution, gene expression, and protein‒protein interaction analyses identify candidate CBL-CIPK signalling networks implicated in stress responses to cold and bacterial infection in citrus.

BACKGROUND: Cold is a major abiotic stress and Huanglongbing and citrus canker disease are two devastating bacterial diseases for citrus. The Ca2+-CBL-CIPK network is known to regulate different types of stress signalling in plants. How do CBL-CIPK signalling networks function in response to cold and infection by CLas or Xcc in citrus? RESULTS: Eight calcineurin B-like proteins (CBLs) and seventeen CBL-interacting protein kinases (CIPKs) were identified from the cold-tolerant satsuma mandarin 'Guijing2501' (Citrus. unshiu) and CLas/Xcc-sensitive sweet orange (C. sinensis). Phylogenetic analysis revealed that both CBL and CIPK family members in citrus were classified into an ancient and a recent clade according to their conserved domain characteristics and/or intron/exon structures. Genome duplication analysis suggested that both tandem and segmental duplications contributed to the amplification of the CBL and CIPK gene families in citrus under intense purifying selection, and the duplication events only existed in the recent clades. Expression comparison of the duplicated gene pairs indicated that the duplicated CBL and CIPK genes underwent functional differentiation. Further expression analysis identified that CBL1, 5, 6, and 8 and CIPK2, 8, 12, 15, 16, and 17 were significantly regulated by multiple stresses, including cold, Xcc infection and/or CLas infection, in citrus, whereas CBL2/7 and CIPK1/4/5/11/13/14 were independently highly regulated by cold and CIPK3 was uniquely responsive to Xcc infection. The combination analyses of targeted Y2H assay and expression analysis revealed that CBL6-CIPK8 was the common signalling network in response to cold and Xcc infection, while CBL6/CBL8-CIPK14 was uniquely responsive to cold in citrus. Further stable transformation and cold tolerance assay indicated that overexpression of CuCIPK16 enhanced the cold tolerance of transgenic Arabidopsis with higher POD activity and lower MDA content. CONCLUSIONS: In this study, evolution, gene expression and protein‒protein interaction analyses of citrus CBLs and CIPKs were comprehensively conducted over a genome-wide range. The results will facilitate future functional characterization of individual citrus CBLs and CIPKs under specific stresses and provide clues for the clarification of cold tolerance and disease susceptibility mechanisms in corresponding citrus cultivars.

Monitor for lactate in perspiration.

Sweat is a noninvasive biological fluid on the surface of human skin and has attracted increasing attention as a diagnostic specimen for disease and biomarker detection. Sweat metabolite quantification is possible due to progress in sweat analysis techniques; nevertheless, the role of sweat monitoring in energy metabolism, physiological or pathological state assessment, health status assessment, and the development and outcome of metabolism-related diseases remains unclear. This review provides a comprehensive overview of the literature on human sweat lactate concentration. The first, second, and third sections of this review present an introduction of sweat lactate, methods for the collection and storage of sweat lactate samples, and methods of detection and analysis of sweat lactate, respectively. The fourth section elaborates upon the current state of clinical application of sweat lactate monitoring and its prospects for health surveillance. The last section focuses on the challenges and future directions of this novel technology for detecting lactate in sweat.

Application of generated mask method based on Mask R-CNN in classification and detection of melanoma.

OBJECTIVE: Melanoma is a type of malignant skin cancer with high mortality, and its incidence is increasing rapidly in recent years. At present, the best treatment is surgical resection after early diagnosis. However, due to the high visual similarity between melanoma and benign melanocytic nevus, coupled with the scarcity and imbalance of data, traditional methods are difficult to achieve good recognition and detection results. Similarly, many machine learning methods have been applied to the task of skin disease detection and classification. However, the accuracy and sensitivity of the experiments are still not satisfactory. Therefore, this paper proposed a method to identify melanoma more efficiently and accurately. METHOD: We implemented a Mixed Skin Lesion Picture Generate method based on Mask R-CNN (MSLP-MR) to solve the problem of data imbalance. Besides, we designed a melanoma detection framework of Mask-DenseNet+ based on MSLP-MR. This method used Mask R-CNN to introduce the method of mask segmentation, and combined with the idea of ensemble learning to integrate multiple classifiers for weighted prediction. Compared with the ablation experiments, the accuracy, sensitivity and AUC of the proposed network classification are improved by 2.56%, 29.33% and 0.0345. RESULT: The experimental results on the ISIC dataset shown that the accuracy of the algorithm is 90.61%, the sensitivity reaches 78.00%, which is higher than the original methods; the specificity reaches 93.43%; and the AUC reaches 0.9502. CONCLUSION: The method is feasible and effective, and achieves the preliminary goal of melanoma detection. It is greatly improved the detection accuracy and reached the level of visual diagnosis of doctors.

Stent placement combined with intraluminal radiofrequency ablation and hepatic arterial infusion chemotherapy for advanced biliary tract cancers with biliary obstruction: a multicentre, retrospective, controlled study.

OBJECTIVE: To evaluate the efficacy and safety of stent placement combined with intraluminal radiofrequency ablation (intra-RFA) and hepatic arterial infusion chemotherapy (HAIC) for patients with advanced biliary tract cancers (Ad-BTCs) and biliary obstruction (BO). METHODS: We retrospectively reviewed data for patients with Ad-BTCs and BO who underwent stent placement with or without intra-RFA and HAIC in three centres between November 2013 and November 2018. The stent patency time (SPT), overall survival (OS), and adverse events (AEs) were analysed. RESULTS: Of the 135 enrolled patients, 64 underwent stent placement combined with intra-RFA and HAIC, while 71 underwent only stent placement. The median SPT was significantly longer in the combination group (8.2 months, 95% confidence interval [CI]: 7.1-9.3) than in the control group (4.3 months, 95% CI: 3.6-5.0; p < 0.001). A similar result was observed for OS (combination: 13.2 months, 95% CI: 11.1-16.5; control: 8.5 months, 95% CI: 7.6-9.6; p < 0.001). The incidence of AEs related to biliary tract operation was not significantly different between the two groups (p > 0.05). The most common AE and serious AE related to HAIC were alanine aminotransferase elevation (24/64; 37.5%) and thrombocytopenia (8/64; 12.5%), respectively. All AEs were tolerable, and there was no death from AEs. CONCLUSIONS: Stent placement combined with intra-RFA and HAIC may be a safe, potential treatment strategy for patients with Ad-BTCs and BO. KEY POINTS: • Advanced biliary cancers (Ad-BTCs) with biliary obstruction (BO) can rapidly result in liver failure and cachexia with an extremely poor prognosis. • Stent placement combined with intraluminal radiofrequency ablation and hepatic arterial infusion chemotherapy may be safe and effective for patients with Ad-BTCs and BO. • The long-term efficacy and safety of the combined treatment is promising.

Tumour stem cell markers CD133 and CD44 are useful prognostic factors after surgical resection of pancreatic neuroendocrine tumours.

The aim of the present study was to investigate the expression profiles and prognostic values of CD133 and CD44 in a cohort of patients with pancreatic neuroendocrine tumours (PNETs). PNET data from patients who underwent radical resection at the Guangdong Provincial People's Hospital were retrospectively analysed. Immunohistochemistry was performed on PNET samples, and CD133 and CD44 expression was examined. Survival analysis was performed using the Kaplan-Meier method and the log-rank test. A total of 71 cases were included in the study. The mean age of the patients was 45.2 years, and the mean tumour size was 3.3 cm. CD44 expression was positively associated with poor tumour differentiation (P=0.007), high Ki-67 index (P=0.001), added mitotic count (P=0.003), high histological grade (P=0.001) and advanced stage (P=0.025). Similarly, CD133 expression was positively associated with high Ki-67 index (P=0.014) and added mitotic count (P=0.012). However, CD133 expression was not associated with tumour differentiation (P=0.118), histological grade (P=0.126) and stage (P=0.203). Survival analysis revealed that both CD44 and CD133 were prognostic factors for overall survival (OS) and/or disease-free survival (DFS), and that increased co-expression of CD44 and CD133 indicated poor OS and DFS rates in patients with PNET. In patients with no expression or low expression of either CD44 or CD133, a DFS rate of 100% was observed, indicating a low recurrence risk. The present findings suggested that high CD44 and CD133 expression was associated with a poor prognosis in patients with PNET. CD44 and CD133 may be used as prognostic indicators of OS and/or DFS in patients with PNETs.

Clinical implication of prognostic and predictive biomarkers for castration-resistant prostate cancer: a systematic review.

BACKGROUND: Diagnosis of metastatic castrate resistant prostate cancer (mCRPC) with current biomarkers is difficult and often results in unnecessary invasive procedures as well as over-diagnosis and over-treatment. There are a number of prognostic biomarkers for CRPC, but there are no validated predictive biomarkers to guide in clinical decision-making. Specific biomarkers are needed that enable to understand the natural history and complex biology of this heterogeneous malignancy, identify early response to treatment outcomes and to identify the population of men most likely to benefit from the treatment. In this systematic review, we discuss the existing literature for the role of biomarkers in CRPC and how they aid in the prognosis, treatment selection and survival outcomes. METHODS: We performed a literature search on PubMed and EMBASE databases from January 2015 through February 2020 in accordance to Preferred Reporting Items for Systematic Review and Meta-Analysis guidelines. Articles were assessed to identify relevant observational studies and randomized controlled trials regarding biomarkers which aid in identifying progression to mCRPC as well as predictive biomarkers which help in treatment selection. RESULTS: We identified 3640 number of hits of which 58 articles were found to be relevant. Here we addressed biomarkers in the context of prognosis, prediction and patient selection of therapy. These biomarkers were found to be effective as prognostic or predictive factors under variety of conditions. The higher levels for all these biomarkers were associated with shorter median OS and sometimes PFS. Lower amounts of biomarkers in serum or urine were associated with prolonged survival outcomes, longer time to CRPC development or CRPC progression and longer median follow-up irrespective of any therapy. CONCLUSION: We observed that the biomarkers included in our study predicted clinically relevant survival outcomes and treatment exposure. Though the current biomarkers are prognostic when measured prior to initiating treatment, not all are validated as predictive markers in post treatment setting. A greater understanding of biomarkers in CRPC is need of the hour for development of more personalized approach to maximize benefit and minimize harm in men with CRPC.

Pancreatic neuroendocrine tumors: A review of serum biomarkers, staging, and management.

Pancreatic neuroendocrine tumors (pNETs) are a heterogeneous group of tumors with complicated treatment options that depend on pathological grading, clinical staging, and presence of symptoms related to hormonal secretion. With regard to diagnosis, remarkable advances have been made: Chromogranin A is recommended as a general marker for pNETs. But other new biomarker modalities, like circulating tumor cells, multiple transcript analysis, microRNA profile, and cytokines, should be clarified in future investigations before clinical application. Therefore, the currently available serum biomarkers are insufficient for diagnosis, but reasonably acceptable in evaluating the prognosis of and response to treatments during follow-up of pNETs. Surgical resection is still the only curative therapeutic option for localized pNETs. However, a debulking operation has also been proven to be effective for controlling the disease. As for drug therapy, steroids and somatostatin analogues are the first-line therapy for those with positive expression of somatostatin receptor, while everolimus and sunitinib represent important progress for the treatment of patients with advanced pNETs. Great progress has been achieved in the combination of systematic therapy with local control treatments. The optimal timing of local control intervention, planning of sequential therapies, and implementation of multidisciplinary care remain pending.

Establishment of a 7-miRNA-Based Risk Score System for Predicting Prognosis of Pancreatic Cancer.

OBJECTIVE: This study developed a prognosis-associated miRNA (PAM)-based risk score system to predict overall survival for pancreatic cancer. METHODS: We screened potential PAMs using bioinformatics technology. A risk score system integrating the PAMs was established, and the predictive value was evaluated. The targets of these PAMs were identified and functional enrichment analysis was performed. RESULTS: Seven PAMs (hsa-mir-188, hsa-mir-1301, hsa-mir-424, hsa-mir-5010, hsa-mir-584, hsa-mir-5091, and hsa-mir-3613) were identified. We also developed a risk score system, which showed a high Harrell concordance index (C-index, 0.723) for overall survival in the Cancer Genome Atlas data sets. The areas under the curve of the receiver operating characteristic curve at the 1-, 2-, and 3-year survival points were 0.718, 0.832, and 0.903, respectively. In addition, both the C-index and the areas under the curve for recurrence-free survival showed a good outcome, indicating that the system had a satisfactory predictive power. Furthermore, 49 target genes of PAMs were identified. Functional enrichment analysis revealed that these targets may be involved in various biological pathways, including the transforming growth factor ß signaling pathway, Notch signaling, and downregulation of SMAD2/3. CONCLUSIONS: The findings of this study suggest that the 7-miRNA-based risk score system is a promising prognostic model for pancreatic cancer.

Potential Mechanisms of AtNPR1 Mediated Resistance against Huanglongbing (HLB) in Citrus.

Huanglongbing (HLB), a bacterial disease caused by Candidatus Liberibacter asiaticus (CLas), is a major threat to the citrus industry. In a previous study conducted by our laboratory, several citrus transgenic trees expressing the Arabidopsis thaliana NPR1 (AtNPR1) gene remained HLB-free when grown in a field site under high HLB disease pressure. To determine the molecular mechanisms behind AtNPR1-mediated tolerance to HLB, a transcriptome analysis was performed using AtNPR1 overexpressing transgenic trees and non-transgenic trees as control, from which we identified 57 differentially expressed genes (DEGs). Data mining revealed the enhanced transcription of genes encoding pathogen-associated molecular patterns (PAMPs), transcription factors, leucine-rich repeat receptor kinases (LRR-RKs), and putative ankyrin repeat-containing proteins. These proteins were highly upregulated in the AtNPR1 transgenic line compared to the control plant. Furthermore, analysis of protein-protein interactions indicated that AtNPR1 interacts with CsNPR3 and CsTGA5 in the nucleus. Our results suggest that AtNPR1 positively regulates the innate defense mechanisms in citrus thereby boosting resistance and effectively protecting the plant against HLB.

Obg-like ATPase 1 (OLA1) overexpression predicts poor prognosis and promotes tumor progression by regulating P21/CDK2 in hepatocellular carcinoma.

BACKGROUND: Obg-like ATPase 1 (OLA1) has been found to have a dual role in cancers. However, the relationship between OLA1 and hepatocellular carcinoma (HCC) remains unclear. RESULTS: High expression of OLA1 in HCC was detected in public datasets and clinical samples, and correlated with poor prognosis. Downregulation of OLA1 significantly inhibited the proliferation, migration, invasion and tumorigenicity of HCC cells. Mechanistically, GSEA showed that OLA1 might promote tumor progression by regulating the cell cycle and apoptosis. In addition, OLA1 knockdown resulted in G0/G1 phase arrest and high levels of apoptosis. OLA1 could bind with P21 and upregulate CDK2 expression to promote HCC progression. CONCLUSIONS: Overall, these findings uncover a role for OLA1 in regulating the proliferation and apoptosis of HCC cells. MATERIALS AND METHODS: The Cancer Genome Atlas and Gene Expression Omnibus datasets were analyzed to identify gene expression. Immunohistochemistry staining, western blot and real-time polymerase chain reaction were performed to evaluate OLA1 expression in samples. Cell count Kit-8, wound-healing, transwell and flow cytometry assays were used to analyze HCC cell progression. Subcutaneous xenotransplantation models were used to investigate the role of OLA1 in vivo. Coimmunoprecipitation was used to analyze protein interactions.

Competitive adsorption of naphthalene and phenanthrene on walnut shell based activated carbon and the verification via theoretical calculation.

Walnut shell based activated carbon (WAC) was prepared via microwave-assisted KOH activation. The adsorption behaviors towards naphthalene (NAP) and phenanthrene (PHE) over WAC were studied, both in single- and binary-compound systems. Characterization results reveal the excellent microporous structure of WAC, with a micropore specific surface area of 438.5 m2 g-1. The functional groups of walnut shell precursor surface were activated through microwave irradiation. In both systems, the pseudo-second-order model can better describe the adsorption kinetic data of PAHs over WAC at all experimental conditions. Mass transfer mechanism analysis shows that film diffusion was the rate-limiting step during the adsorption process. The adsorption amount of PAHs on WAC decreased as pH values increased, and the equilibrium data can be fitted by the Freundlich isotherm model well. In binary-component systems, the presence of PHE prominently restrained the adsorption towards NAP, and the Sheindorf-Rebhun-Sheintuch (SRS) model can fully fit the adsorption equilibrium experimental values of PAHs over WAC. In addition, the preferential adsorption behavior of PHE over WAC also was confirmed by theoretical calculations. The π-π complex between the active sites on the WAC surface and π-electrons of benzene rings from PAHs may play a major role in competitive adsorption. These results indicated that WAC was a potentially low-cost adsorbent for PAH elimination.

Selenium concentration, dietary intake and risk of hepatocellular carcinoma - A systematic review with meta-analysis / Concentración de selenio, ingesta dietética y riesgo de carcinoma hepatocelular: revisión sistemática con metaanálisis

Aim: this study was performed to investigate the association between selenium concentrations, dietary intake, and the risk of hepatocellular carcinoma (HCC). Methods: we identified eligible studies in PubMed and EMBASE databases, in addition to the reference lists of original studies and review articles on this topic, up to 1 Feb 2019. A summary of standardized mean differences (SMD) with 95% confidence intervals (CI) was calculated using a random-effects model. Heterogeneity between studies was assessed using Cochran Q and I2 statistics. Results: finally, a meta-analysis showed that dietary intake of selenium and tissue selenium concentration were not associated with HCC risk (dietary SMD = -0.11, 95% CI: -0.26 to 0.03; tissue SMD = -0.12, 95% CI: -0.56 to 0.33). However, samples from toenail, whole blood, and serum all showed an inverse association with HCC risk (toenail SMD = -0.53, 95% CI: -0.72 to -0.35; whole blood SMD = -2.21, 95% CI: -2.67 to -1.76; tissue SMD = -1.26, 95% CI: -1.71 to -0.81). Dose-response data from few studies showed that an extra increase in serum selenium was dramatically related with a lower risk of HCC (adjusted p-trend < 0.05). This study showed that selenium concentration in toenail, whole blood and serum was inversely associated with HCC risk. Conclusion: increased concentration in serum selenium was related to a lower risk of HCC. However, these results based on dietary intake and tissue samples, which included few studies, did not reach statistical significance

Objetivo: este estudio se realizó para investigar la asociación entre las concentraciones de selenio, la ingesta dietética y el riesgo de carcinoma hepatocelular (CHC). Métodos: identificamos estudios elegibles en las bases de datos PubMed y EMBASE, además de las listas de referencias de los estudios originales y artículos de revisión sobre este tema hasta el 1 de febrero de 2019. Se realizó un resumen de las diferencias medias estandarizadas (SMD) con intervalos de confianza (CI) del 95% utilizando un modelo de efectos aleatorios. La heterogeneidad entre estudios se evaluó utilizando las estadísticas de Cochran Q e I2. Resultados: por último, el metaanálisis mostró que la concentración de selenio en la ingesta dietética y de selenio tisular no estaban asociadas al riesgo de HCC (SMD dietética -0,11, IC 95%: -0,26 a 0,03; SMD tisular -0,12, IC 95%: -0,56 a 0,33). Sin embargo, las muestras de uña del pie, sangre entera y suero mostraron todas ellas una asociación inversa con el riesgo de CHC (SMD ungueal -0.53, IC 95%: -0.72 a -0.35; SMD de sangre entera -2.21, IC 95%: -2.67 a -1.76; SMD tisular -1.26, IC 95%: -1.71 a -0.81). Los datos de dosis-respuesta de pocos estudios mostraron que los incrementos del selenio sérico se relacionaban fuertemente con un menor riesgo de CHC (tendencia de p ajustada < 0.05). Este estudio demostró que la concentración de selenio en las uñas del pie, en sangre entera y en suero se asocian inversamente al riesgo de CHC. Conclusión: el aumento de la concentración de selenio sérico se relacionó con menor riesgo de CHC. Sin embargo, los resultados de la ingesta dietética y los tejidos, que incluían pocos estudios, no alcanzaron la significación estadística

Selenium concentration, dietary intake and risk of hepatocellular carcinoma - A systematic review with meta-analysis.

INTRODUCTION: Aim: this study was performed to investigate the association between selenium concentrations, dietary intake, and the risk of hepatocellular carcinoma (HCC). Methods: we identified eligible studies in PubMed and EMBASE databases, in addition to the reference lists of original studies and review articles on this topic, up to 1 Feb 2019. A summary of standardized mean differences (SMD) with 95% confidence intervals (CI) was calculated using a random-effects model. Heterogeneity between studies was assessed using Cochran Q and I2 statistics. Results: finally, a meta-analysis showed that dietary intake of selenium and tissue selenium concentration were not associated with HCC risk (dietary SMD = -0.11, 95% CI: -0.26 to 0.03; tissue SMD = -0.12, 95% CI: -0.56 to 0.33). However, samples from toenail, whole blood, and serum all showed an inverse association with HCC risk (toenail SMD = -0.53, 95% CI: -0.72 to -0.35; whole blood SMD = -2.21, 95% CI: -2.67 to -1.76; tissue SMD = -1.26, 95% CI: -1.71 to -0.81). Dose-response data from few studies showed that an extra increase in serum selenium was dramatically related with a lower risk of HCC (adjusted p-trend < 0.05). This study showed that selenium concentration in toenail, whole blood and serum was inversely associated with HCC risk. Conclusion: increased concentration in serum selenium was related to a lower risk of HCC. However, these results based on dietary intake and tissue samples, which included few studies, did not reach statistical significance.

INTRODUCCIÓN: Objetivo: este estudio se realizó para investigar la asociación entre las concentraciones de selenio, la ingesta dietética y el riesgo de carcinoma hepatocelular (CHC). Métodos: identificamos estudios elegibles en las bases de datos PubMed y EMBASE, además de las listas de referencias de los estudios originales y artículos de revisión sobre este tema hasta el 1 de febrero de 2019. Se realizó un resumen de las diferencias medias estandarizadas (SMD) con intervalos de confianza (CI) del 95% utilizando un modelo de efectos aleatorios. La heterogeneidad entre estudios se evaluó utilizando las estadísticas de Cochran Q e I2. Resultados: por último, el metaanálisis mostró que la concentración de selenio en la ingesta dietética y de selenio tisular no estaban asociadas al riesgo de HCC (SMD dietética -0,11, IC 95%: -0,26 a 0,03; SMD tisular -0,12, IC 95%: -0,56 a 0,33). Sin embargo, las muestras de uña del pie, sangre entera y suero mostraron todas ellas una asociación inversa con el riesgo de CHC (SMD ungueal -0.53, IC 95%: -0.72 a -0.35; SMD de sangre entera -2.21, IC 95%: -2.67 a -1.76; SMD tisular -1.26, IC 95%: -1.71 a -0.81). Los datos de dosis-respuesta de pocos estudios mostraron que los incrementos del selenio sérico se relacionaban fuertemente con un menor riesgo de CHC (tendencia de p ajustada < 0.05). Este estudio demostró que la concentración de selenio en las uñas del pie, en sangre entera y en suero se asocian inversamente al riesgo de CHC. Conclusión: El aumento de la concentración de selenio sérico se relacionó con menor riesgo de CHC. Sin embargo, los resultados de la ingesta dietética y los tejidos, que incluían pocos estudios, no alcanzaron la significación estadística.

Comparative safety and effectiveness of ultrasound-guided radiofrequency ablation combined with preoperative three-dimensional reconstruction versus surgical resection for solitary hepatocellular carcinoma of 3-5 cm.

OBJECTIVE: To investigate the safety and effectiveness of ultrasound-guided radiofrequency ablation (RFA) combined with preoperative three-dimensional (3D) reconstruction versus surgical resection for solitary hepatocellular carcinoma of 3-5 cm. METHODS: The cohort of this retrospective study included 66 consecutive patients who underwent open hepatectomy (Surgery group) between January 2009 and December 2014, as well as 54 consecutive patients who underwent ultrasound-guided RFA combined with preoperative 3D reconstruction (RFA group) during the same period. Preoperative 3D reconstruction was performed using Myrian-XP-Liver software. The image fusion system was used to evaluate the RFA safety margin at 1 month after surgery. Kaplan-Meier analysis and the log-rank test were used to compare the recurrence and overall survival (OS) rates between the two treatment groups. RESULTS: There were no significant differences in the baseline characteristics of the two groups. The complete ablation rate was 94.4% (51/53). As compared with surgical resection for solitary HCC of 3-5 cm, ultrasound-guided RFA combined with preoperative 3D reconstruction significantly reduced the morbidity of excessive pain, total complications, and infections (p < 0.001). A significant decrease in the duration of the hospital stay after treatment was also observed in the RFA group (t = 10.017, p < 0.001). There was no significant difference in the cumulative recurrence rate between the two groups. Kaplan-Meier analysis and the log-rank test revealed no significant difference in the OS rate between the two groups over a 3-year follow-up period. CONCLUSION: Ultrasound-guided RFA combined with preoperative 3D reconstruction appears to be a safe and effective therapeutic option for patients with solitary HCC of 3-5 cm.

Decreased expression of LKB1 predicts poor prognosis in pancreatic neuroendocrine tumor patients undergoing curative resection.

BACKGROUND: Liver kinase B1 (LKB1) is a key regulatory protein of cellular metabolism, proliferation, and polarity. The present study aimed to characterize the expression pattern of LKB1 in pancreatic neuroendocrine tumors (pNETs) and evaluate the relationship between LKB1 expression and prognosis in pNETs. PATIENTS AND METHODS: We retrospectively analyzed the pathologic and clinical data of 71 pNET patients who underwent curative surgical resection in Guangdong General Hospital. LKB1 mRNA and protein levels in tumor tissues and paired nontumor tissues were evaluated in 24 patients by quantitative real-time reverse-transcription polymerase chain reaction and Western blot, respectively. Immunohistochemical expression of LKB1 in tumor tissues was detected in all of the 71 patients, and the immunohistochemical expression level was re-coded in two classes (high versus low/negative) and correlated with clinicopathological parameters and survival outcomes. The association between LKB1 expression and clinicopathological characters was evaluated by chi-square test and Student's t-test. Kaplan-Meier curves and log-rank test were used to analyze the survival outcomes, including overall survival (OS) and disease-free survival (DFS). RESULTS: Compared to adjacent normal tissues, LKB1 mRNA level and protein expression level in tumor tissues were both increased. The immunostaining of LKB1 was mainly found within the cytoplasm. Overall, 52 of 71 (73.2%) cases were positive for LKB1 protein, which showed either a diffuse staining pattern or a partial staining pattern. Decreased LKB1 expression was correlated with older age (P=0.042), increased Ki-67 index (P=0.004), increased mitotic count (P=0.001), and advanced histologic grade (P=0.001). Moreover, patients with low/negative LKB1 expression had shorter OS and DFS than those with high expression. CONCLUSION: Our results suggested that LKB1 expression could be a useful prognostic marker for recurrence and survival in pNET patients who had received curative resection.

Evaluation of the three-in-one team-based care model on hierarchical diagnosis and treatment patterns among patients with diabetes: a retrospective cohort study using Xiamen's regional electronic health records.

BACKGROUND: Xiamen is a pilot city in China for hierarchical diagnosis and treatment reform of non-communicable diseases, especially diabetes. Since 2012, Xiamen has implemented a program called the "three-in-one", a team-based care model for the treatment of diabetes, which involves collaboration between diabetes specialists, general practitioners, and health managers. In addition, the program provides financial incentives to improve care, as greater accessibility to medications through community health care centers (CHCs). The aim of this study was to evaluate the effectiveness of these policies in shifting visits from general hospitals to CHCs for the treatment of type 2 diabetes mellitus (T2DM). METHOD AND MATERIALS: A retrospective observational cohort study was conducted using Xiamen's regional electronic health record (EHR) database, which included 90% of all patients registered since 2012. Logistic regression was used to derive the adjusted odds ratio (OR) for patients shifting from general hospitals to CHCs. Among patients treated at hospitals, Kaplan-Meier(KM) curves were constructed to evaluate the time from each policy introduction until the switch to CHCs. A k-means clustering analysis was conducted to identify patterns of patient care-seeking behavior. RESULTS: In total, 89,558 patients and 2,373,524 visits were included. In contrast to increased outpatient visits to general hospitals in China overall, the percentage of visits to CHCs in Xiamen increased from 29.7% in 2012 to 66.5% in 2016. The most significant and rapid shift occurred in later periods after full policy implementation. Three clusters of patients were identified with different levels of complications and health care-seeking frequency. All had similar responses to the policies. CONCLUSIONS: The "three-in-one" team-based care model showed promising results for building a hierarchical health care system in China. These policy reforms effectively increased CHCs utilization among diabetic patients.

Candidate Resistant Genes of Sand Pear (Pyrus pyrifolia Nakai) to Alternaria alternata Revealed by Transcriptome Sequencing.

Pear black spot (PBS) disease, which is caused by Alternaria alternata (Aa), is one of the most serious diseases affecting sand pear (Pyrus pyrifolia Nakai) cultivation worldwide. To investigate the defense mechanisms of sand pear in response to Aa, the transcriptome of a sand pear germplasm with differential resistance to Aa was analyzed using Illumina paired-end sequencing. Four libraries derived from PBS-resistant and PBS-susceptible sand pear leaves were characterized through inoculation or mock-inoculation. In total, 20.5 Gbp of sequence data and 101,632,565 reads were generated, representing 44717 genes. Approximately 66% of the genes or sequenced reads could be aligned to the pear reference genome. A large number (5213) of differentially expressed genes related to PBS resistance were obtained; 34 microsatellites were detected in these genes, and 28 genes were found to be closely related to PBS resistance. Using a transcriptome analysis in response to PBS inoculation and comparison analysis to the PHI database, 4 genes (Pbr039001, Pbr001627, Pbr025080 and Pbr023112) were considered to be promising candidates for sand pear resistance to PBS. This study provides insight into changes in the transcriptome of sand pear in response to PBS infection, and the findings have improved our understanding of the resistance mechanism of sand pear to PBS and will facilitate future gene discovery and functional genome studies of sand pear.

[Effects of different calcium salts on freezing resistance of satsuma mandarin].

The study on the effects of foliar spraying 20 mmol x L(-1) of CaCl2, Ca (NO3)2 and CaAc2 on the freezing resistance of satsuma mandarin (Citrus unshiu Marc. cv. Guoqing No. 1) showed that after treated with these Ca salts, the leaves of test plant had a lower half lethal temperature (LT50) than the control (sprayed with distilled water). The LT50 after treated with CaCl2 was 0.54 degrees C lower, whereas that after treated with CaAc2 and Ca (NO3)2 was 1.34 degrees C and 1.35 degrees C lower, respectively, implying that the latter two Ca salts were more effective in enhancing the freezing resistance of satsuma mandarin. Moreover, foliar spraying Ca salts increased the superoxidase (SOD) and peroxidase (POD) activities and the contents of soluble proteins, soluble sugars and praline in leaves, and decreased the leaf MDA content.